Protein kinase modulators

ABSTRACT

The invention relates in part to molecules having certain biological activities that include, but are not limited to, inhibiting cell proliferation, modulating protein kinase activity and modulating polymerase activity. Molecules of the invention can modulate Pim kinase activity and/or FMS-like tyrosine kinase (Flt) activity. The invention also relates in part to methods for using such molecules.

RELATED APPLICATIONS

This application claims benefit of priority to U.S. ProvisionalApplication Ser. No. 61/067,845, filed 29 Feb. 2008, and U.S.Provisional Application Ser. No. 61/103,908, filed 8 Oct. 2008, thecontents of each of which are incorporated herein by reference in theirentirety.

FIELD OF THE INVENTION

The invention relates in part to molecules having certain biologicalactivities that include, but are not limited to, inhibiting cellproliferation, modulating serine-threonine protein kinase activity andmodulating tyrosine kinase activity. Molecules of the invention canmodulate casein kinase (CK) activity (e.g., CK2 activity), Pim kinaseactivity (e.g., PIM-1, PIM-2 and/or PIM-3 activity) and/or Fms-liketyrosine kinase (Flt) activity (e.g., Flt-3 activity). The inventionalso relates in part to methods for using such molecules.

BACKGROUND ART

The PIM protein kinases which include the closely related PIM-1, -2, and-3, have been implicated in diverse biological processes such as cellsurvival, proliferation, and differentiation. PIM-1 is involved in anumber of signaling pathways that are highly relevant to tumorigenesis[reviewed in Bachmann & Moroy, Internat. J. Biochem. Cell Biol., 37,726-730 (2005)]. Many of these are involved in cell cycle progressionand apoptosis. It has been shown that PIM-1 acts as an anti-apoptoticfactor via inactivation of the pro-apoptotic factor BAD (Bcl2 associateddeath promoter, an apoptosis initiator). This finding suggested a directrole of PIM-1 in preventing cell death, since the inactivation of BADcan enhance Bcl-2 activity and can thereby promote cell survival [Aho etal., FEBS Letters, 571, 43-49 (2004)]. PIM-1 has also been recognized asa positive regulator of cell cycle progression. PIM-1 binds andphosphorylates Cdc25A, which leads to an increase in its phosphataseactivity and promotion of G1/S transition [reviewed in Losman et al.,JBC, 278, 4800-4805 (1999)]. In addition, the cyclin kinase inhibitorp21^(Waf) which inhibits G1/S progression, was found to be inactivatedby PIM-1 [Wang et al., Biochim. Biophys. Act. 1593, 45-55 (2002)].Furthermore, by means of phosphorylation, PIM-1 inactivates C-TAKl andactivates Cdc25C which results in acceleration of G2/M transition[Bachman et al., JBC, 279, 48319-48 (2004)].

PIM-1 appears to be an essential player in hematopoietic proliferation.Kinase active PIM-1 is required for the gp130-mediated STAT3proliferation signal [Hirano et al., Oncogene 19, 2548-2556, (2000)].PIM-1 is overexpressed or even mutated in a number of tumors anddifferent types of tumor cell lines and leads to genomic instability.Fedorov, et al., concluded that a Phase III compound in development fortreating leukemia, LY333′531, is a selective PIM-1 inhibitor. O.Fedorov, et al., PNAS 104(51), 20523-28 (December 2007). Evidence hasbeen published to show that PIM-1 is involved in human tumors includingprostate cancer, oral cancer, and Burkitt lymphoma (Gaidano & DallaFaver, 1993). All these findings point to an important role of PIM-1 inthe initiation and progression of human cancers, including varioustumors and hematopoietic cancers, thus small molecule inhibitors ofPIM-1 activity are a promising therapeutic strategy.

Additionally, PIM-2 and PIM-3 have overlapping functions with PIM-1 andinhibition of more than one isoform may provide additional therapeuticbenefits. However, it is sometimes preferable for inhibitors of PIM tohave little or no in vivo impact through their inhibition of variousother kinases, since such effects are likely to cause side effects orunpredictable results. See, e.g., O. Fedorov, et al., PNAS 104(51),20523-28 (December 2007), discussing the effects that non-specifickinase inhibitors can produce. Accordingly, in some embodiments, theinvention provides compounds that are selective inhibitors of at leastone of PIM-1, PIM-2, and PIM-3, or some combination of these, whilehaving substantially less activity on certain other human kinases, asdescribed further herein.

The implication of a role for PIM-3 in cancer was first suggested bytranscriptional profiling experiments showing that PIM3 genetranscription was upregulated in EWS/ETS-induced malignanttransformation of NIH 3T3 cells. These results were extended to showthat PIM-3 is selectively expressed in human and mouse hepatocellularand pancreatic carcinomas but not in normal liver or pancreatic tissues.In addition, PIM-3 mRNA and protein are constitutively expressed inmultiple human pancreatic and hepatocellular cancer cell lines.

The link between PIM-3 overexpression and a functional role in promotingtumorigenesis came from RNAi studies in human pancreatic andhepatocellular cancer cell lines overexpressing PIM-3. In these studiesthe ablation of endogenous PIM-3 protein promoted apoptosis of thesecells. The molecular mechanism by which PIM-3 suppresses apoptosis is inpart carried out through the modulation of phosphorylation of thepro-apoptotic protein BAD. Similar to both PIM-1 & 2 which phosphorylateBAD protein, the knockdown of PIM-3 protein by siRNA results in adecrease in BAD phosphorylation at Ser112. Thus, similar to PIM-1 and 2,PIM-3 acts a suppressor of apoptosis in cancers of endodermal origin,e.g., pancreatic and liver cancers. Moreover, as conventional therapiesin pancreatic cancer have a poor clinical outcome, PIM-3 could representa new important molecular target towards successful control of thisincurable disease.

At the 2008 AACR Annual Meeting, SuperGen announced that it hasidentified a lead PIM kinase inhibitor, SGI-1776, that causes tumorregression in acute myelogenous leukemia (AML) xenograft models(Abstract No. 4974). In an oral presentation entitled, “A potent smallmolecule PIM kinase inhibitor with activity in cell lines fromhematological and solid malignancies,” Dr. Steven Warner detailed howscientists used SuperGen's CLIMB™ technology to build a model thatallowed for the creation of small molecule PIM kinase inhibitors.SGI-1776 was identified as a potent and selective inhibitor of the PIMkinases, inducing apoptosis and cell cycle arrest, thereby causing areduction in phospho-BAD levels and enhancement of mTOR inhibition invitro. Most notably, SGI-1776 induced significant tumor regression inMV-4-11 (AML) and MOLM-13 (AML) xenograft models. This demonstrates thatinhibitors of PIM kinases can be used to treat leukemias.

Fedorov, et al., in PNAS vol. 104(51), 20523-28, showed that a selectiveinhibitor of PIM-1 kinase (Ly5333′531) suppressed cell growth andinduced cell death in leukemic cells from AML patients. PIM-3 has beenshown to be expressed in pancreatic cancer cells, while it is notexpressed in normal pancreas cells, demonstrating that it should be agood target for pancreatic cancer. Li, et al., Cancer Res. 66(13),6741-47 (2006).

Another kinase shown to be a useful target for certain cancers,including leukemia, is Flt3 kinase (FMS-like tyrosine kinase 3). Flt3 isprevalent in refractory AML patients, so inhibitors of Flt3 are usefulto treat such patients. Smith, et al., reported an alkaloid calledCEP-701 that is a potent inhibitor of Flt3 and provided clinicalresponses in tested subjects with minimal dose-related toxicity. Blood,vol. 103(10), 3669-76 (2004). Dual inhibitors that are active againstboth PIM and Flt3 may be advantageous over inhibitors of either targetalone. In particular, excessive Flt3 activity is associated withrefractory AML, so dual inhibitors of PIM and Flt3 such as compoundsdisclosed herein are useful to treat refractory AML.

In addition, Flt3 inhibitors are useful to treat inflammation.Inhibitors of Flt3 have been shown to be effective to treat airwayinflammation in mice, using a murine asthma model. Edwan, et al., J.Immunologoy, 5016-23 (2004). Accordingly, the compounds of theinvention, are useful to treat conditions associated with excessiveactivity of Flt3, including inflammation such as airway inflammation andasthma.

Collectively, these results demonstrate that inhibitors of PIM kinasesand Flt3 kinase are useful for treating certain types of cancers.Accordingly, the identification of compounds that specifically inhibit,regulate and/or modulate the signal transduction of PIM-1, PIM-2, PIM-3,and/or Flt3 is desirable as a means to treat or prevent disease statesassociated with abnormal cell proliferation, such as cancer. Theinvention provides compounds, compositions and methods that address thisneed and are useful for treating cancers, inflammation and pain.

DISCLOSURE OF THE INVENTION

The present invention in part provides chemical compounds having certainbiological activities that include, but are not limited to, inhibitingcell proliferation, inhibiting angiogenesis, and modulating protein kinase activity. These molecules can modulate casein kin ase 2 (CK2)activity, Pim kinase activity, and/or Fms-like tyrosine kinase 3 (Flt)activity and thus affect biological functions that include but are notlimited to, inhibiting gamma phosphate transfer from ATP to a protein orpeptide substrate, inhibiting angiogenesis, inhibiting cellproliferation and inducing cell apoptosis, for example. The presentinvention also in part provides methods for preparing novel chemicalcompounds, and analogs thereof, and methods of using the foregoing. Alsoprovided are compositions comprising the above-described molecules incombination with other agents, and methods for using such molecules incombination with other agents.

The compounds of the invention have the general formula (A):

wherein the group labeled α represents a 5-6 membered aromatic orheteroaromatic ring fused onto the ring containing Q¹, wherein α is a6-membered aryl ring optionally containing one or more nitrogen atoms asring members, or a five membered aryl ring selected from thiophene andthiazole;

Q¹ is C═X, Q² is NR⁵, and the bond between Q¹ and Q² is a single bond;or Q¹ is C—X—R⁵, Q² is N, and the bond between Q¹ and Q² is a doublebond; and

wherein X represents O, S or NR⁴, and Z¹-Z⁸ and R⁴ and R⁵ are as definedbelow;

provided that when Q¹ in Formula (A) is C—NHΦ, where Φ is optionallysubstituted phenyl:

if the ring labeled α is a six-membered ring containing at least one Nas a ring member, at least one R³ present must be a polar substituent,or if each R³ is H, then Φ must be substituted; and

if the ring labeled α is phenyl, and three of Z¹-Z⁴ represent CH, thenZ² cannot be C—OR″, and Z³ cannot be NH₂, NO₂, NHC(═O)R″ or NHC(═O)—OR″,where R″ is C1-C4 alkyl.

The invention also includes the pharmaceutically acceptable salts ofcompounds of formula (A). Thus in each compound of the invention,Formula (A) represents a fused tricyclic ring system which is linkedthrough either Q1 or Q2 to a group R5, which is further described below.

Thus, provided herein are compounds of Formulae I, II, III and IV:

and pharmaceutically acceptable salts, esters, prodrugs and tautomersthereof; wherein:

each Z¹, Z², Z³, and Z⁴ is N or CR³;

each of Z⁵, Z⁶, Z⁷ and Z⁸ is CR⁶ or N;

each R³ and each R⁶ is independently H or an optionally substitutedC1-C8 alkyl, C2-C8 heteroalkyl, C2-C8 alkenyl, C2-C8 heteroalkenyl,C2-C8 alkynyl, C2-C8 heteroalkynyl, C1-C8 acyl, C2-C8 heteroacyl, C6-C10aryl, C5-C12 heteroaryl, C7-C12 arylalkyl, or C6-C12 heteroarylalkylgroup,

or each R³ and each R⁶ can be halo, OR, NR₂, NROR, NRNR₂, SR, SOR, SO₂R,SO₂NR₂, NRSO₂R, NRCONR₂, NRCSNR₂, NRC(═NR)NR₂, NRCOOR, NRCOR, CN, COOR,CONR₂, OOCR, COR, polar substituent, carboxy bioisostere, or NO₂,

wherein each R is independently H or C1-C8 alkyl, C2-C8 heteroalkyl,C2-C8 alkenyl, C2-C8 heteroalkenyl, C2-C8 alkynyl, C2-C8 heteroalkynyl,C1-C8 acyl, C2-C8 heteroacyl, C6-C10 aryl, C5-C10 heteroaryl, C7-C12arylalkyl, or C6-C12 heteroarylalkyl,

and wherein two R on the same atom or on adjacent atoms can be linked toform a 3-8 membered ring, optionally containing one or more N, O or S;

and each R group, and each ring formed by linking two R groups together,is optionally substituted with one or more substituents selected fromhalo, ═O, ═N—CN, ═N—OR′, ═NR′, OR′, NR′₂, SR′, SO₂R′, SO₂NR′₂, NR′SO₂R′,NR′CONR′₂, NR′CSNR′₂, NR′C(═NR′)NR′₂, NR′COOR′, NR′COR′, CN, COOR′,CONR′₂, OOCR′, COR′, and NO₂,

wherein each R′ is independently H, C1-C6 alkyl, C2-C6 heteroalkyl,C1-C6 acyl, C2-C6 heteroacyl, C6-C10 aryl, C5-C10 heteroaryl, C7-12arylalkyl, or C6-12 heteroarylalkyl, each of which is optionallysubstituted with one or more groups selected from halo, C1-C4 alkyl,C1-C4 heteroalkyl, C1-C6 acyl, C1-C6 heteroacyl, hydroxy, amino, and ═O;

and wherein two R′ can be linked to form a 3-7 membered ring optionallycontaining up to three heteroatoms selected from N, O and S,

R⁴ is H or optionally substituted member selected from the groupconsisting of C₁-C₆ alkyl, C2-C6 heteroalkyl, and C1-C6 acyl;

each R⁵ is independently H or an optionally substituted member selectedfrom the group consisting of C₁₋₁₀ alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀heteroalkyl, C₃₋₈ carbocyclic ring, and C₃₋₈ heterocyclic ringoptionally fused to an additional optionally substituted carbocyclic orheterocyclic; or R⁵ is a C₁₋₁₀ alkyl, C₂₋₁₀ alkenyl, or C₂₋₁₀heteroalkyl substituted with an optionally substituted C₃₋₈ carbocyclicring or C₃₋₈ heterocyclic ring; and

in each —NR⁴R⁵, R⁴ and R⁵ together with N may form an optionallysubstituted 3-8 membered ring, which may optionally contain anadditional heteroatom selected from N, O and S as a ring member;

provided that when —NR⁴R⁵ in Formula (I) is —NHD, where Φ is optionallysubstituted phenyl:

if at least one of Z⁵-Z⁸ is N, at least one R³ present must be a polarsubstituent, or if each R³ is H, then Φ must be substituted; and

if each of Z⁵-Z⁸ is CR⁶, and three of Z¹-Z⁴ represent CH, then Z² cannotbe C—OR″, and Z³ cannot be NH₂, NO₂, NHC(═O)R″ or NHC(═O)—OR″, where R″is C1-C4 alkyl.

In certain embodiments, provided are compounds having the structure ofFormulae I, II, III, and IV, and pharmaceutically acceptable salts,esters and tautomers thereof; wherein:

each Z¹, Z², Z³, and Z⁴ is N or CR³;

each of Z⁵, Z⁶, Z⁷ and Z⁸ is N or CR⁶;

none, one or two of Z¹-Z⁴ are N and none, one or two of Z⁵-Z⁸ are N;

each R³ and each R⁶ is independently H or an optionally substitutedC1-C8 alkyl, C2-C8 heteroalkyl, C2-C8 alkenyl, C2-C8 heteroalkenyl,C2-C8 alkynyl, C2-C8 heteroalkynyl, C1-C8 acyl, C2-C8 heteroacyl, C6-C10aryl, C5-C12 heteroaryl, C7-C12 arylalkyl, or C6-C12 heteroarylalkylgroup,

or each R³ and each R⁶ is independently halo, OR, NR₂, NROR, NRNR₂, SR,SOR, SO₂R, SO₂NR₂, NRSO₂R, NRCONR₂, NRCSNR₂, NRC(═NR)NR₂, NRCOOR, NRCOR,CN, COOR, CONR₂, OOCR, COR, polar substituent, carboxy bioisostere, orNO₂,

wherein each R is independently H or C1-C8 alkyl, C2-C8 heteroalkyl,C2-C8 alkenyl, C2-C8 heteroalkenyl, C2-C8 alkynyl, C2-C8 heteroalkynyl,C1-C8 acyl, C2-C8 heteroacyl, C6-C10 aryl, C5-C10 heteroaryl, C7-C12arylalkyl, or C6-C12 heteroarylalkyl,

and wherein two R on the same atom or on adjacent atoms can be linked toform a 3-8 membered ring, optionally containing one or more N, O or S;

and each R group, and each ring formed by linking two R groups together,is optionally substituted with one or more substituents selected fromhalo, ═O, ═N—CN, ═N—OR′, ═NR′, OR′, NR′₂, SR′, SO₂R′, SO₂NR′₂,NR′SO₂R′,NR′CONR′₂, NR′CSNR′₂, NR′C(═NR′)NR′₂, NR′COOR′, NR′COR′, CN,COOR′, CONR′₂, OOCR′, COR′, and NO₂,

wherein each R′ is independently H, C1-C6 alkyl, C2-C6 heteroalkyl,C1-C6 acyl, C2-C6 heteroacyl, C6-C10 aryl, C5-C10 heteroaryl, C7-12arylalkyl, or C6-12 heteroarylalkyl, each of which is optionallysubstituted with one or more groups selected from halo, C1-C4 alkyl,C1-C4 heteroalkyl, C1-C6 acyl, C1-C6 heteroacyl, hydroxy, amino, and ═O;

and wherein two R′ can be linked to form a 3-7 membered ring optionallycontaining up to three heteroatoms selected from N, O and S;

R⁴ is H or an optionally substituted member selected from the groupconsisting of C1-C6 alkyl, C2-C6 heteroalkyl, and C1-C6 acyl;

each R⁵ is independently H or an optionally substituted member selectedfrom the group consisting of C₁₋₁₀ alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀heteroalkyl, C₃₋₈ carbocyclic ring, and C₃₋₈ heterocyclic ringoptionally fused to an additional optionally substituted carbocyclic orheterocyclic; or R⁵ is a C₁₋₁₀ alkyl, C₂₋₁₀ alkenyl, or C₂₋₁₀heteroalkyl substituted with an optionally substituted C₃₋₈ carbocyclicring or C₃₋₈ heterocyclic ring; and in each —NR⁴R⁵, R⁴ and R⁵ togetherwith N may form an optionally substituted 3-8 membered ring, which mayoptionally contain an additional heteroatom selected from N, O and S asa ring member;

provided that when —NR⁴R⁵ in Formula (I) is —NHΦ, where Φ is optionallysubstituted phenyl:

if all of Z⁵-Z⁸ are CH or one of Z⁵-Z⁸ is N, at least one of Z¹-Z⁴ isCR³ and at least one R³ must be a non-hydrogen substituent; or

if each R³ is H, then Φ must be substituted; or

if all of Z⁵-Z⁸ are CH or one of Z⁵-Z⁸ is N, then Z² is not C—OR″, andZ³ is not NH₂, NO₂, NHC(═O)R″ or NHC(═O)—OR″, where R″ is C1-C4 alkyl.

In certain embodiments of Formulae I, II, III, and IV, one, two, threeor four of Z⁵, Z⁶, Z⁷ and Z⁸ are N. For embodiments in which two of Z⁵,Z⁶, Z⁷ and Z⁸ are N, the ring nitrogen atoms may be adjacent (e.g.,nitrogen atoms at Z⁵ and Z⁶, Z⁶ and Z⁷, or Z⁷ and Z⁸) or may beseparated by one or two ring positions (e.g., nitrogen atoms at Z⁵ andZ⁷, Z⁶ and Z⁸ or Z⁵ and Z⁸). In frequent embodiments, at least one R³substituent is a polar substituent, such as a carboxylic acid or a salt,an ester or a bioisostere thereof. In some embodiments, at least one R³is a carboxylic acid-containing substituent or a carboxylatebioisostere, or a salt or ester thereof, for example. In someembodiments, at least one R³ is a carboxylic acid-containing substituentor a salt thereof. In certain embodiments, at least one R³ is acarboxamide. In other embodiments, at least one R³ is a C₁₋₃ alkylsubstituted with SO₂NR₂, NRSO₂R, NRCONR₂, NRCSNR₂, NRC(═NR)NR₂, NRCOOR,NRCOR, or CONR₂, wherein each R is independently H or C1-C8 alkyl, C2-C8heteroalkyl, C2-C8 alkenyl, C2-C8 heteroalkenyl, C2-C8 alkynyl, C2-C8heteroalkynyl, C1-C8 acyl, C2-C8 heteroacyl, C6-C10 aryl, C5-C10heteroaryl, C7-C12 arylalkyl, or C6-C12 heteroarylalkyl.

The term “polar substituent” as used herein refers to any substituenthaving an electric dipole, and optionally a dipole moment (e.g., anasymmetrical polar substituent has a dipole moment and a symmetricalpolar substituent does not have a dipole moment). Polar substituentsinclude substituents that accept or donate a hydrogen bond, and groupsthat would carry at least a partial positive or negative charge inaqueous solution at physiological pH levels. In certain embodiments, apolar substituent is one that can accept or donate electrons in anon-covalent hydrogen bond with another chemical moiety. In certainembodiments, a polar substituent is selected from a carboxy, a carboxybioisostere or other acid-derived moiety that exists predominately as ananion at a pH of about 7 to 8. Other polar substituents include, but arenot limited to, groups containing an OH or NH, an ether oxygen, an aminenitrogen, an oxidized sulfur or nitrogen, a carbonyl, a nitrile, and anitrogen-containing or oxygen-containing heterocyclic ring whetheraromatic or non-aromatic. In some embodiments, the polar substituentrepresented by R³ is a carboxylate or a carboxylate bioisostere.

“Carboxylate bioisostere” or “carboxy bioisostere” as used herein refersto a moiety that is expected to be negatively charged to a substantialdegree at physiological pH. In certain embodiments, the carboxylatebioisostere is a moiety selected from the group consisting of:

and salts and prodrugs of the foregoing, wherein each R⁷ isindependently H or an optionally substituted member selected from thegroup consisting of C₁₋₁₀ alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ heteroalkyl, C₃₋₈carbocyclic ring, and C₃₋₈ heterocyclic ring optionally fused to anadditional optionally substituted carbocyclic or heterocyclic ring; orR⁷ is a C₁₋₁₀ alkyl, C₂₋₁₀ alkenyl, or C₂₋₁₀ heteroalkyl substitutedwith an optionally substituted C₃₋₈ carbocyclic ring or C₃₋₈heterocyclic ring. In certain embodiments, the polar substituent (e.g.,R^(3P)) is selected from the group consisting of carboxylic acid,carboxylic ester, carboxamide, tetrazole, triazole, imidazole,carboxymethanesulfonamide, oxadiazole, oxothiadiazole, thiazole,aminothiazole and hydroxythiazole. In some embodiments, at least one R³present is a carboxylic acid or a salt, or ester or a bioisosterethereof. In certain embodiments, at least one R³ present is a carboxylicacid-containing substituent or a salt, ester or bioisostere thereof. Inthe latter embodiments, the R³ substituent may be a C1-C10 alkyl orC1-C10 alkenyl linked to a carboxylic acid (or salt, ester orbioisostere thereof), for example, and in some embodiments, the R³substituent is not —NHCOOCH₂CH₃.

In some preferred embodiments of the present invention, R^(3P) is atriazole or imidazole ring, which can be substituted or unsubstituted,and is preferably bonded through a carbon atom of the triazole orimidazole ring to the fused tricyclic moiety. R^(3P) is frequently a2-imidazolyl ring or a 3-triazolyl ring, each of which can beunsubstituted or substituted. If these rings are substituted on N, theyare typically substituted with C1-C6 alkyl or C1-C6 acyl, or, ifsubstituted on a carbon atom of the ring, with halo. Unsubstituted3-triazole is a preferred group for R^(3P).

In certain embodiments, at least one of Z¹-Z⁴ and Z⁵-Z⁸ is a nitrogenatom, and one or more ring nitrogen atoms can be positioned in the ringcontaining Z¹-Z⁴ or in the ring containing Z⁵-Z⁸ such that each ring isindependently an optionally substituted pyridine, pyrimidine, pyridazineor pyrazine ring. For example, one or more ring nitrogen atoms withinthe ring containing Z⁵-Z⁸ may be arranged as follows:

where each R^(6A), R^(6B), R^(6C) and R^(6D) independently is selectedfrom R⁶ substituents defined above with respect to compounds of FormulaI, II, III or IV.

In certain embodiments, no two adjacent Z¹-Z⁴ or Z⁵-Z⁸ both are N.

A polar substituent may be at any position on the ring containing Z¹-Z⁴in Formula I, II, III or IV, and the ring may include one, two, three orfour polar substituents. In certain embodiments, each of Z¹-Z⁴ may beCR³ and one of the R³ substituents may be a polar substituent (e.g., acarboxylate or carboxylic acid ester, carboxamide or a tetrazole)arranged at any one of the positions in the ring containing Z¹-Z⁴:

where R^(3P) is a polar substituent and each R^(3A), R^(3B), R^(3C) andR^(3D) independently is selected from R³ substituents, as defined abovewith respect to compounds of Formula I, II, III or IV.

In certain embodiments of the compounds in the foregoing Formulae, R⁴ isH. In some embodiments, R⁴ is H or CH₃ and R⁵ is an optionallysubstituted 3-8 membered ring, which can be aromatic, nonaromatic, andcarbocyclic or heterocyclic, or R⁵ is a C₁₋₁₀ alkyl group substitutedwith such an optionally substituted 3-8 membered ring. In specificembodiments, R⁵ is an optionally substituted five-, six-, orseven-membered carbocyclic or heterocyclic ring, and sometimes is anoptionally substituted phenyl ring.

In some embodiments pertaining to compounds of Formula I, R⁴ is H or CH₃and R⁵ is a phenyl substituted with one or more halogen (e.g., F, Cl),fluoroalkyl (e.g., CF₃) or acetylene substituents, which substituentssometimes are on the phenyl ring at the 3-position, 4-position or5-position, or combinations thereof (e.g., the 3- and 5-positions).

R⁵ in certain embodiments is a C₁₋₃ alkyl substituted with an optionallysubstituted phenyl, pyridyl, morpholino, piperidinyl or pyrrolidinylring substituent, or is substituted with hydroxyl or —NR⁴R⁴ where R⁴ isas defined above (e.g., R⁵ may be C₁₋₃ alkyl substituted with —N(CH₃)₂).In other embodiments, R⁵ is a C₁₋₃ alkyl substituted with SO₂NR₂,NRSO₂R, NRCONR₂, NRCOOR, NRCOR, or CONR₂, wherein each R isindependently H or C1-C8 alkyl, C2-C8 heteroalkyl, C2-C8 alkenyl, C2-C8heteroalkenyl, C2-C8 alkynyl, C2-C8 heteroalkynyl, C1-C8 acyl, C2-C8heteroacyl, C6-C10 aryl, C5-C10 heteroaryl, C7-C12 arylalkyl, or C6-C12heteroarylalkyl. The polar group represented by R³ in some embodimentsis a carboxy, carboxyalkyl (e.g., carboxymethyl), tetrazole orcarboxamide (e.g., —CONH₂) substituent. In other embodiments, R³represents a carboxylate bioisostere.

An R⁶ substituent in certain embodiments, such as R^(6B), sometimes is a—NR⁴R⁵ substituent, such as a —NH—(C₁-C₆ alkyl) moiety (e.g., —NH—CH₃),for example. In some embodiments, the compound has the structure ofFormula I; R⁴ is H or CH₃; R⁵ is an optionally substituted five-, six-,or seven-membered carbocyclic or heterocyclic ring, and sometimes is anoptionally substituted phenyl ring; and one R³ is a carboxylic acid or asalt, an ester, carboxamide or a carboxylate bioisostere. In someembodiments, the compound has the structure of Formula I; R⁴ is H orCH₃; R⁵ is an optionally substituted five-, six-, or seven-memberedcarbocyclic or heterocyclic ring, and sometimes is an optionallysubstituted phenyl ring; and one or two of Z⁵, Z⁶, Z⁷ and Z⁸ are N.

In some embodiments of compounds of Formulae I, II, III or IV, each ofZ¹, Z², Z³, and Z⁴ is CR³, and at least one R³ is H, or at least two R³are H. Often, at least one R⁶ is H, or at least two R⁶ are H. In someembodiments, (i) each Z¹, Z², Z³, Z⁴, Z⁵, Z⁶ and Z⁸ is CR³ and Z⁷ isnitrogen; or (ii) each Z¹, Z², Z³, Z⁴, Z⁶, Z⁷ and Z⁸ is CR³ and Z⁵ isnitrogen; or (iii) each Z¹, Z², Z³, Z⁴, Z⁶, and Z⁸ is CR³ and each of Z⁵and Z⁷ is nitrogen. Each R³ and/or each R⁶ present in certainembodiments is hydrogen, except that at least one R³ present is a polarsubstituent. In some embodiments, each R^(3A), R^(3C), R^(3D), R^(6A),R^(6B), R^(6C) and R^(6D) is H and R^(3B) is a polar substituent (e.g.,carboxylate, carboxylic acid, tetrazole).

Also provided herein are compounds of Formulae V, VI, VII or VIII:

and pharmaceutically acceptable salts, esters, prodrugs and tautomersthereof; where Z¹, Z², Z³, Z⁴, R⁴ and R⁵ are defined above with respectto compounds of Formulae I, II, III and IV, and each R^(6A) and R^(6B)is independently selected from an R⁶ substituent defined above withrespect to compounds of Formulae I, II, III and IV.

As with compounds of Formulae I, II, III and IV, in preferredembodiments at least one R³ present is a polar substituent, such as apolar substituent described above. In frequent embodiments, at least oneR³ substituent is a polar substituent, such as a carboxylic acid or asalt, an ester or a bioisostere thereof. In some embodiments, at leastone R³ is a carboxylic acid-containing substituent or a carboxylatebioisostere, or a salt or ester thereof, for example. In someembodiments, at least one R³ is a carboxylic acid-containing substituentor a salt thereof. In other embodiments, at least one R³ is acarboxamide. Embodiments described with respect to compounds of FormulaeI, II, III and IV also may be applied to compounds of Formulae V, VI,VII and VIII.

In certain embodiments, provided are compounds having a structure ofFormulae V, VI, VII and VIII, and pharmaceutically acceptable salts,esters and tautomers thereof; wherein:

each Z¹, Z², Z³, and Z⁴ independently is N or CR³ and none, one or twoof Z¹, Z², Z³, and Z⁴ is N;

each R³, R^(6A) and R^(6B) independently is H or an optionallysubstituted C1-C8 alkyl, C2-C8 heteroalkyl, C2-C8 alkenyl, C2-C8heteroalkenyl, C2-C8 alkynyl, C2-C8 heteroalkynyl, C1-C8 acyl, C2-C8heteroacyl, C6-C10 aryl, C5-C12 heteroaryl, C7-C12 arylalkyl, or C6-C12heteroarylalkyl group,

or each R³, R^(6A) and R^(6B) independently is halo, OR, NR₂, NROR,NRNR₂, SR, SOR, SO₂R, SO₂NR₂, NRSO₂R, NRCONR₂, NRCSNR₂, NRC(═NR)NR₂,NRCOOR, NRCOR, CN, COOR, polar substituent, carboxy bioisostere, CONR₂,OOCR, COR, or NO₂,

wherein each R is independently H or C1-C8 alkyl, C2-C8 heteroalkyl,C2-C8 alkenyl, C2-C8 heteroalkenyl, C2-C8 alkynyl, C2-C8 heteroalkynyl,C1-C8 acyl, C2-C8 heteroacyl, C6-C10 aryl, C5-C10 heteroaryl, C7-C12arylalkyl, or C6-C12 heteroarylalkyl,

and wherein two R on the same atom or on adjacent atoms can be linked toform a 3-8 membered ring, optionally containing one or more N, O or S;

and each R group, and each ring formed by linking two R groups together,is optionally substituted with one or more substituents selected fromhalo, ═O, ═N—CN, ═N—OR′, ═NR′, OR′, NR′₂, SR′, SO₂R′, SO₂NR′₂, NR′SO₂R′,NR′CONR′₂, NR′CSNR′₂, NR′C(═NR′)NR′₂, NR′COOR′, NR′COR′, CN, COOR′,CONR′₂, OOCR′, COR′, and NO₂,

wherein each R′ is independently H, C1-C6 alkyl, C2-C6 heteroalkyl,C1-C6 acyl, C2-C6 heteroacyl, C6-C10 aryl, C5-C10 heteroaryl, C7-12arylalkyl, or C6-12 heteroarylalkyl, each of which is optionallysubstituted with one or more groups selected from halo, C1-C4 alkyl,C1-C4 heteroalkyl, C1-C6 acyl, C1-C6 heteroacyl, hydroxy, amino, and ═O;

and wherein two R′ can be linked to form a 3-7 membered ring optionallycontaining up to three heteroatoms selected from N, O and S,

R⁴ is H or optionally substituted member selected from the groupconsisting of C₁-C₆ alkyl, C₂-C₆ heteroalkyl, and C1-C6 acyl;

each R⁵ is independently H or an optionally substituted member selectedfrom the group consisting of C₁₋₁₀ alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀heteroalkyl, C₃₋₈ carbocyclic ring, and C₃₋₈ heterocyclic ringoptionally fused to an additional optionally substituted carbocyclic orheterocyclic; or R⁵ is a C₁₋₁₀ alkyl, C₂₋₁₀ alkenyl, or C₂₋₁₀heteroalkyl substituted with an optionally substituted C₃₋₈ carbocyclicring or C₃₋₈ heterocyclic ring; and

in each —NR⁴R⁵, R⁴ and R⁵ together with N may form an optionallysubstituted 3-8 membered ring, which may optionally contain anadditional heteroatom selected from N, O and S as a ring member.

In some embodiments pertaining to compounds of Formulae V, VI, VII andVIII, each of Z¹, Z², Z³, and Z⁴ is CR³, and at least one R³ is H, or atleast two R³ are H. Often, at least one of R^(6A) and R^(6B) is H, andsometimes each of R^(6A) and R^(6B) is H. In certain embodiments, eachR³ and/or each of R^(6A) and R^(6B) present is H, except that at leastone R³ present is a polar substituent. In some embodiments, each R^(3A),R^(3C), R^(3D), R^(6A) and R^(6B) is H and R^(3B) is a polar substituent(e.g., carboxylate bioisostere, carboxylic acid, carboxamide ortetrazole).

In certain embodiments pertaining to compounds of Formula V, R⁴ is H orCH₃ and R⁵ is an optionally substituted five-, six- or seven-memberedcarbocyclic or heterocyclic ring (e.g., optionally substituted phenylring). In some embodiments pertaining to compounds of Formula V, R⁴ is Hor CH₃ and R⁵ is a phenyl ring substituted with one or more halogen(e.g., F, Cl), trifluoroalkyl (e.g., CF₃), or acetylene substituents,which substituents sometimes are at the 3-position, 4-position or5-position, or a combination thereof (e.g., the 3- and 5-positions). R⁵in certain embodiments is a C₁₋₃ alkyl substituted with an optionallysubstituted phenyl, pyridyl, morpholino, pyrrolyl, piperidinyl orpyrrolidinyl substituent, or a C₁₋₃ alkyl substituted with a hydroxyl orwith a substituent —NR⁴R⁴, where R⁴ is as defined above (e.g., R⁵ can beC₁₋₃ alkyl substituted with —N(CH₃)_(z)). In other embodiments, R⁵ is aC₁₋₃ alkyl substituted with SO₂NR₂, NRSO₂R, NRCONR₂, NRCSNR₂,NRC(═NR)NR₂, NRCOOR, NRCOR, or CONR₂, wherein each R is independently Hor C1-C8 alkyl, C2-C8 heteroalkyl, C2-C8 alkenyl, C2-C8 heteroalkenyl,C2-C8 alkynyl, C2-C8 heteroalkynyl, C1-C8 acyl, C2-C8 heteroacyl, C6-C10aryl, C5-C10 heteroaryl, C7-C12 arylalkyl, or C6-C12 heteroarylalkyl. AnR⁶ substituent in certain embodiments, such as R^(6A) or R^(6B),sometimes is a —NR⁴R⁵ substituent, such as a —NH—(C₁-C₆ alkyl) moiety(e.g., —NH—CH₃), for example. In other embodiments, each of R^(6A) andR^(6B) is H.

Provided also are compounds of Formulae IX, X, XI and XII:

and pharmaceutically acceptable salts, esters, prodrugs and tautomersthereof; where Z¹, Z², Z³, Z⁴, R⁴, R⁵ and R⁶ are defined above withrespect to compounds of Formulae I, II, III and IV.

As with compounds of Formulae I, II, III and IV, in frequentembodiments, at least one R³ present is a polar substituent, such as apolar substituent described above (e.g., carboxylic acid, carboxylate,carboxamide tetrazole). For compounds of Formula IX, R⁴ and R⁵ are notboth hydrogen, and independently are H, —Y⁰ or -LY¹, where Y⁰ is anoptionally substituted 5-membered ring or optionally substituted6-membered ring (e.g., heterocyclic ring or carbocyclic ring each beingaryl or non-aryl), Y¹ is an optionally substituted 5-membered aryl ringor optionally substituted 6-membered aryl ring, and L is a C1-C20 alkyllinker or C1-C20 alkylene linker.

In some embodiments, provided are compounds having a structure ofFormulae IX, X, XI and XII, and pharmaceutically acceptable salts,esters and tautomers thereof; wherein:

each Z¹, Z², Z³, and Z⁴ is N or CR³ and none, one or two of Z¹, Z², Z³,and Z⁴ is N;

each R³ and R⁶ is independently H or an optionally substituted C₁-C8alkyl, C2-C8 heteroalkyl, C2-C8 alkenyl, C2-C8 heteroalkenyl, C2-C8alkynyl, C2-C8 heteroalkynyl, C1-C8 acyl, C2-C8 heteroacyl, C6-C10 aryl,C5-C12 heteroaryl, C7-C12 arylalkyl, or C6-C12 heteroarylalkyl group,

or each R³ and R⁶ can be halo, OR, NR₂, NROR, NRNR₂, SR, SOR, SO₂R,SO₂NR₂, NRSO₂R, NRCONR₂, NRCSNR₂, NRC(═NR)NR₂, NRCOOR, NRCOR, CN, COOR,polar substituent, carboxy bioisostere, CONR₂, OOCR, COR, or NO₂,

wherein each R is independently H or C1-C8 alkyl, C2-C8 heteroalkyl,C2-C8 alkenyl, C2-C8 heteroalkenyl, C2-C8 alkynyl, C2-C8 heteroalkynyl,C1-C8 acyl, C2-C8 heteroacyl, C6-C10 aryl, C5-C10 heteroaryl, C7-C12arylalkyl, or C6-C12 heteroarylalkyl,

and wherein two R on the same atom or on adjacent atoms can be linked toform a 3-8 membered ring, optionally containing one or more N, O or S;

and each R group, and each ring formed by linking two R groups together,is optionally substituted with one or more substituents selected fromhalo, ═O, ═N—CN, ═N—OR′, ═NR′, OR′, NR′₂, SR′, SO₂R′, SO₂NR′₂, NR′SO₂R′,NR′CONR′₂, NR′CSNR′₂, NR′C(═NR′)NR′₂, NR′COOR′, NR′COR′, CN, COOR′,CONR′₂, OOCR′, COR′, and NO₂,

wherein each R′ is independently H, —C6 alkyl, C2-C6 heteroalkyl, C1-C6acyl, C2-C6 heteroacyl, C6-C10 aryl, C5-C10 heteroaryl, C7-12 arylalkyl,or C6-12 heteroarylalkyl, each of which is optionally substituted withone or more groups selected from halo, C1-C4 alkyl, C1-C4 heteroalkyl,C1-C6 acyl, C1-C6 heteroacyl, hydroxy, amino, and ═O;

and wherein two R′ can be linked to form a 3-7 membered ring optionallycontaining up to three heteroatoms selected from N, O and S;

R⁴ is H or optionally substituted member selected from the groupconsisting of C₁-C₆ alkyl, C2-C6 heteroalkyl, and C1-C6 acyl;

each R⁵ is independently H or an optionally substituted member selectedfrom the group consisting of C₁₋₁₀ alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀heteroalkyl, C₃₋₈ carbocyclic ring, and C₃₋₈ heterocyclic ringoptionally fused to an additional optionally substituted carbocyclic orheterocyclic; or R⁵ is a C₁₋₁₀ alkyl, C₂₋₁₀ alkenyl, or C₂₋₁₀heteroalkyl substituted with an optionally substituted C₃₋₈ carbocyclicring or C₃₋₈ heterocyclic ring; and

in each —NR⁴R⁵, R⁴ and R⁵ together with N may form an optionallysubstituted 3-8 membered ring, which may optionally contain anadditional heteroatom selected from N, O and S as a ring member.

Embodiments described with respect to compounds of Formulae I, II, III,IV, V, VI, VII and VIII also may be applied to compounds of Formulae IX,X, XI and XII. In some embodiments pertaining to compounds of FormulaeIX, X, XI and XII, each of Z¹, Z², Z³, and Z⁴ is CR³, and at least oneR³ is H, or at least two R³ are H. R⁶ often is H, and in certainembodiments, each R⁶ and R³ present is H, except that at least one R³present is a polar substituent. In some embodiments, each R^(3A),R^(3C), R^(3D) and R⁶ is H and R^(3B) is a polar substituent (e.g.,carboxylate, carboxylic acid, carboxamide, or tetrazole).

In certain embodiments pertaining to compounds of Formula IX, R⁴ is H orCH₃ and R⁵ is an optionally substituted five-, six- or seven-memberedcarbocyclic or heterocyclic ring (e.g., optionally substituted phenylring). In some embodiments pertaining to compounds of Formula IX, R⁴ isH or CH₃ and R⁵ is a phenyl ring substituted with one or more halogen(e.g., F, Cl) or acetylene substituents, which substituents sometimesare at the 3-position, 4-position or 5-position, or a combinationthereof (e.g., the 3- and 5-positions). R⁵ in certain embodiments is aC₁₋₃ alkyl substituted with an optionally substituted phenyl, pyridyl,morpholino, pyrrolyl, piperidinyl or pyrrolidinyl substituent, or a C₁₋₃alkyl substituted with a hydroxyl substituent or substituted with a—NR⁴R⁴ (e.g., —N(CH₃)₂) substituent. In other embodiments, R⁵ is a C₁₋₃alkyl substituted with SO₂NR₂, NRSO₂R, NRCONR₂, NRCSNR_(Z), NRC(═NR)NR₂,NRCOOR, NRCOR, or CONR₂, wherein each R is independently H or C1-C8alkyl, C2-C8 heteroalkyl, C2-C8 alkenyl, C2-C8 heteroalkenyl, C2-C8alkynyl, C2-C8 heteroalkynyl, C1-C8 acyl, C2-C8 heteroacyl, C6-C10 aryl,C5-C10 heteroaryl, C7-C12 arylalkyl, or C6-C12 heteroarylalkyl. R⁶ incertain embodiments sometimes is a —NR⁴R⁵ substituent, such as a—NH—(C1-C6 alkyl) moiety (e.g., —NH—CH₃), for example.

Also provided herein are compounds of Formulae XIII, XIV, XV and XVI:

and pharmaceutically acceptable salts, esters, prodrugs and tautomersthereof; wherein:

Z5 is N or CR6A;

each R6A, R6B, R6C and R8 independently is H or an optionallysubstituted C1-C8 alkyl, C2-C8 heteroalkyl, C2-C8 alkenyl, C2-C8heteroalkenyl, C2-C8 alkynyl, C2-C8 heteroalkynyl, C1-C8 acyl, C2-C8heteroacyl, C6-C10 aryl, C5-C12 heteroaryl, C7-C12 arylalkyl, or C6-C12heteroarylalkyl group,

or each R6A, R6B, R6C and R8 independently is halo, CF3, CFN, OR, NR2,NROR, NRNR2, SR, SOR, SO2R, SO2NR2, NRSO2R, NRCONR2, NRCSNR2,NRC(═NR)NR2, NRCOOR, NRCOR, CN, COOR, carboxy bioisostere, CONR2, OOCR,COR, or NO2,

R9 is independently an optionally substituted C1-C8 alkyl, C2-C8heteroalkyl, C2-C8 alkenyl, C2-C8 heteroalkenyl, C2-C8 alkynyl, C2-C8heteroalkynyl, C1-C8 acyl, C2-C8 heteroacyl, C6-C10 aryl, C5-C12heteroaryl, C7-C12 arylalkyl, or C6-C12 heteroarylalkyl group, or

R9 is independently halo, OR, NR2, NROR, NRNR2, SR, SOR, SO2R, SO2NR2,NRSO2R, NRCONR2, NRCOOR, NRCOR, CN, COOR, CONR2, OOCR, COR, or NO2,

wherein each R is independently H or C1-C8 alkyl, C2-C8 heteroalkyl,C2-C8 alkenyl, C2-C8 heteroalkenyl, C2-C8 alkynyl, C2-C8 heteroalkynyl,C1-C8 acyl, C2-C8 heteroacyl, C6-C10 aryl, C5-C10 heteroaryl, C7-C12arylalkyl, or C6-C12 heteroarylalkyl,

and wherein two R on the same atom or on adjacent atoms can be linked toform a 3-8 membered ring, optionally containing one or more N, O or S;

and each R group, and each ring formed by linking two R groups together,is optionally substituted with one or more substituents selected fromhalo, ═O, ═N—CN, ═N—OR′, ═NR′, OR′, NR′2, SR′, SO2R′, SO2NR′2, NR′SO2R′,NR′CONR′2, NR′CSNR′2, NR′C(═NR′)NR′2, NR′COOR′, NR′COR′, CN, COOR′,CONR′2, OOCR′, COR′, and NO2,

wherein each R′ is independently H, C1-C6 alkyl, C2-C6 heteroalkyl,C1-C6 acyl, C2-C6 heteroacyl, C6-C10 aryl, C5-C10 heteroaryl, C7-12arylalkyl, or C6-12 heteroarylalkyl, each of which is optionallysubstituted with one or more groups selected from halo, C1-C4 alkyl,C1-C4 heteroalkyl, C1-C6 acyl, C1-C6 heteroacyl, hydroxy, amino, and ═O;

and wherein two R′ can be linked to form a 3-7 membered ring optionallycontaining up to three heteroatoms selected from N, O and S;

n is 0 to 4; and

p is 0 to 4.

In certain embodiments for compounds of Formulae XIII, XIV, XV and XVI,Z⁵ is N. In some embodiments, R⁸ is a carboxy moiety, such as acarboxylate or carboxylic acid. In certain embodiments, R⁹ is selectedfrom —C≡CR, —C≡CH, —CH₃, —CH₂CH₃, —CF₃, —CFN, —OR or halogen. In someembodiments R⁹ is selected from halogen, —C≡CR or —C≡CH. In certainembodiments R⁹ is selected from halogen or —C≡CH, and in someembodiments R⁹ is halogen, is chloro, is bromo or is —C≡CH

Also provided herein are compounds of Formulae XVII, XVIII, XIX or XX:

and pharmaceutically acceptable salts, esters, prodrugs and tautomersthereof; where Z¹, Z², Z³, Z⁴ and R⁵ are defined above with respect tocompounds of Formulae I, II, III and IV, and each R^(6A) and R^(6B) isindependently selected from an R⁶ substituent defined above with respectto compounds of Formulae I, II, III and IV.

As with compounds of Formulae I, II, III and IV, in frequent embodimentsat least one R3 present is a polar substituent, such as a polarsubstituent described above. Embodiments described with respect tocompounds of Formulae I, II, III and IV also may be applied to compoundsof Formulae XVII, XVIII, XIX or XX.

In certain embodiments, provided are compounds having a structure ofFormulae XVII, XVIII, XIX or XX, and pharmaceutically acceptable salts,esters and tautomers thereof; wherein:

each Z¹, Z², Z³, and Z⁴ independently is N or CR³ and none, one or twoof Z¹, Z², Z³, and Z⁴ is N;

each R³, R^(6A) and R^(6B) independently is H or an optionallysubstituted C1-C8 alkyl, C2-C8 heteroalkyl, C2-C8 alkenyl, C2-C8heteroalkenyl, C2-C8 alkynyl, C2-C8 heteroalkynyl, C1-C8 acyl, C2-C8heteroacyl, C6-C10 aryl, C5-C12 heteroaryl, C7-C12 arylalkyl, or C6-C12heteroarylalkyl group,

or each R³, R^(6A) and R^(6B) independently is halo, OR, NR₂, NROR,NRNR₂, SR, SOR, SO₂R, SO₂NR₂, NRSO₂R, NRCONR₂, NRCSNR₂, NRC(═NR)NR₂,NRCOOR, NRCOR, CN, COOR, polar substituent, carboxy bioisostere, CONR₂,OOCR, COR, or NO₂,

wherein each R is independently H or C1-C8 alkyl, C2-C8 heteroalkyl,C2-C8 alkenyl, C2-C8 heteroalkenyl, C2-C8 LLalkynyl, C2-C8heteroalkynyl, C1-C8 acyl, C2-C8 heteroacyl, C6-C10 aryl, C5-C10heteroaryl, C7-C12 arylalkyl, or C6-C12 heteroarylalkyl,

and wherein two R on the same atom or on adjacent atoms can be linked toform a 3-8 membered ring, optionally containing one or more N, O or S;

and each R group, and each ring formed by linking two R groups together,is optionally substituted with one or more substituents selected fromhalo, ═O, ═N—CN, ═N—OR′, ═NR′, OR′, NR′₂, SR′, SO₂R′, SO₂NR′₂, NR′SO₂R′,NR′CONR′₂, NR′CSNR′₂, NR′C(═NR′)NR′₂, NR′COOR′, NR′COR′, CN, COOR′,CONR′₂, OOCR′, COR′, and NO₂,

wherein each R′ is independently H, C1-C6 alkyl, C2-C6 heteroalkyl,C1-C6 acyl, C2-C6 heteroacyl, C6-C10 aryl, C5-C10 heteroaryl, C7-12arylalkyl, or C6-12 heteroarylalkyl, each of which is optionallysubstituted with one or more groups selected from halo, C1-C4 alkyl,C1-C4 heteroalkyl, C1-C6 acyl, C1-C6 heteroacyl, hydroxy, amino, and ═O;

and wherein two R′ can be linked to form a 3-7 membered ring optionallycontaining up to three heteroatoms selected from N, O and S;

R⁴ is H or optionally substituted member selected from the groupconsisting of C₁-C₆ alkyl, C2-C6 heteroalkyl, and C1-C6 acyl;

each R⁵ is independently H or an optionally substituted member selectedfrom the group consisting of C₁₋₁₀ alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀heteroalkyl, C₃₋₈ carbocyclic ring, and C₃₋₈ heterocyclic ringoptionally fused to an additional optionally substituted carbocyclic orheterocyclic; or R⁵ is a C₁₋₁₀ alkyl, C₂₋₁₀ alkenyl, or C₂₋₁₀heteroalkyl substituted with an optionally substituted C₃₋₈ carbocyclicring or C₃₋₈ heterocyclic ring; and

in each —NR⁴R⁵, R⁴ and R⁵ together with N may form an optionallysubstituted 3-8 membered ring, which may optionally contain anadditional heteroatom selected from N, O and S as a ring member.

In some embodiments pertaining to compounds of Formulae XVII, XVIII, XIXor XX, each of Z¹, Z², Z³, and Z⁴ is CR³, and at least one R³ is H, orat least two R³ are H. Often, at least one of R^(6A) and R^(6B) is H,and sometimes each of R^(6A) and R^(6B) is H. In certain embodiments,each R³ and/or each of R^(6A) and R^(6B) present is H, except that atleast one R³ present is a polar substituent. In some embodiments, eachR^(3A), R^(3C), R^(3D), R^(6A) and R^(6B) is H and R^(3B) is a polarsubstituent (e.g., carboxylate bioisostere, carboxylic acid,carboxamide, or tetrazole).

In certain embodiments pertaining to compounds of Formula XVII, R⁴ is Hor CH₃ and R⁵ is an optionally substituted five-, six- or seven-memberedcarbocyclic or heterocyclic ring (e.g., optionally substituted phenylring). In some embodiments pertaining to compounds of Formula XVII, R⁴is H or CH₃ and R⁵ is a phenyl ring substituted with one or more halogen(e.g., F, Cl), fluoroalkyl (e.g., CF₃) or acetylene substituents, whichsubstituents sometimes are at the 3-position, 4-position or 5-position,or a combination thereof (e.g., the 3- and 5-positions). R⁵ in certainembodiments is a C₁₋₃ alkyl substituted with an optionally substitutedphenyl, pyridyl, morpholino, pyrrolyl, piperidinyl or pyrrolidinylsubstituent, or a C₁₋₃ alkyl substituted with a hydroxyl substituent orsubstituted with a substituent —NR⁴R⁴, where R⁴ is as defined above(e.g., —N(CH₃)₂). In other embodiments, R⁵ is a C₁₋₃ alkyl substitutedwith SO₂NR₂, NRSO₂R, NRCONR₂, NRCSNR₂, NRC(═NR)NR₂, NRCOOR, NRCOR, orCONR₂, wherein each R is independently H or C1-C8 alkyl, C2-C8heteroalkyl, C2-C8 alkenyl, C2-C8 heteroalkenyl, C2-C8 alkynyl, C2-C8heteroalkynyl, C1-C8 acyl, C2-C8 heteroacyl, C6-C10 aryl, C5-C10heteroaryl, C7-C12 arylalkyl, or C6-C12 heteroaryl alkyl. An R⁶substituent in certain embodiments, such as R^(6A) or R^(6B), sometimesis a halo, or —NR⁴R⁵ substituent, such as a —NH—(C1-C6 alkyl) moiety(e.g., —NH—CH₃), for example.

Also provided herein are compounds of Formulae XXI, XXII, XXIII or XXIV:

and pharmaceutically acceptable salts, esters, prodrugs and tautomersthereof; where Z¹, Z², Z³, Z⁴, R⁴ and R⁵ are defined above with respectto compounds of Formulae I, II, III and IV, and each R^(6A) and R^(6B)is independently selected from an R⁶ substituent defined above withrespect to compounds of Formulae I, II, III and IV. As with compounds ofFormulae I, II, III and IV, in frequent embodiments at least one R³present is a polar substituent, such as a polar substituent describedabove. Embodiments described with respect to compounds of Formulae I,II, III and IV also may be applied to compounds of Formulae XXI, XXII,XXIII or XXIV.

In certain embodiments, provided are compounds having a structure ofFormulae XXI, XXII, XXIII or XXIV, and pharmaceutically acceptablesalts, esters and tautomers thereof; wherein:

each Z¹, Z², Z³, and Z⁴ independently is N or CR³ and none, one or twoof Z¹, Z², Z³, and Z⁴ is N;

each R³, R^(6A) and R^(6B) independently is H or an optionallysubstituted C1-C8 alkyl, C2-C8 heteroalkyl, C2-C8 alkenyl, C2-C8heteroalkenyl, C2-C8 alkynyl, C2-C8 heteroalkynyl, C1-C8 acyl, C2-C8heteroacyl, C6-C10 aryl, C5-C12 heteroaryl, C7-C12 arylalkyl, or C6-C12heteroarylalkyl group,

or each R³, R^(6A) and R^(6B) independently is halo, OR, NR₂, NROR,NRNR₂, SR, SOR, SO₂R, SO₂NR₂, NRSO₂R, NRCONR₂, NRCSNR₂, NRC(═NR)NR₂,NRCOOR, NRCOR, CN, COOR, polar substituent, carboxy bioisostere, CONR₂,OOCR, COR, or NO₂,

wherein each R is independently H or C1-C8 alkyl, C2-C8 heteroalkyl,C2-C8 alkenyl, C2-C8 heteroalkenyl, C2-C8 alkynyl, C2-C8 heteroalkynyl,C1-C8 acyl, C2-C8 heteroacyl, C6-C10 aryl, C5-C10 heteroaryl, C7-C12arylalkyl, or C6-C12 heteroarylalkyl,

and wherein two R on the same atom or on adjacent atoms can be linked toform a 3-8 membered ring, optionally containing one or more N, O or S;

and each R group, and each ring formed by linking two R groups together,is optionally substituted with one or more substituents selected fromhalo, ═O, ═N—CN, ═N—OR′, ═NR′, OR′, NR′₂, SR′, SO₂R′, SO₂NR′₂, NR′SO₂R′,NR′CONR′₂, NR′COOR′, NR′CSNR′₂, NR′C(═NR′)NR′₂, NR′COR′, CN, COOR′,CONR′₂, OOCR′, COR′, and NO₂,

wherein each R′ is independently H, C1-C6 alkyl, C2-C6 heteroalkyl,C1-C6 acyl, C2-C6 heteroacyl, C6-C10 aryl, C5-C10 heteroaryl, C7-12arylalkyl, or C6-12 heteroarylalkyl, each of which is optionallysubstituted with one or more groups selected from halo, C1-C4 alkyl,C1-C4 heteroalkyl, C1-C6 acyl, C1-C6 heteroacyl, hydroxy, amino, and ═O;

and wherein two R′ can be linked to form a 3-7 membered ring optionallycontaining up to three heteroatoms selected from N, O and S;

R⁴ is H or optionally substituted member selected from the groupconsisting of C1-C6 alkyl, C2-C6 heteroalkyl, and C1-C6 acyl;

each R⁵ is independently H or an optionally substituted member selectedfrom the group consisting of C₁₋₁₀ alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀heteroalkyl, C₃₋₈ carbocyclic ring, and C₃₋₈ heterocyclic ringoptionally fused to an additional optionally substituted carbocyclic orheterocyclic; or R⁵ is a C₁₋₁₀ alkyl, C₂₋₁₀ alkenyl, or C₂₋₁₀heteroalkyl substituted with an optionally substituted C₃₋₈ carbocyclicring or C₃₋₈ heterocyclic ring; and

in each —NR⁴R⁵, R⁴ and R⁵ together with N may form an optionallysubstituted 3-8 membered ring, which may optionally contain anadditional heteroatom selected from N, O and S as a ring member.

In some embodiments pertaining to compounds of Formulae XXI, XXII, XXIIIor XXIV, each of Z¹, Z², Z³, and Z⁴ is CR³, and at least one R³ is H, orat least two R³ are H. Often, at least one of R^(6A) and R^(6B) is H,and sometimes each of R^(6A) and R^(6B) is H. In certain embodiments,each R³ and/or each of R^(6A) and R^(6B) present is H, except that atleast one R³ present is a polar substituent. In some embodiments, eachR^(3A), R^(3C), R^(3D), R^(6A) and R^(6B) is H and R^(3B) is a polarsubstituent (e.g., carboxylate bioisostere, carboxylic acid,carboxamide, or tetrazole).

In certain embodiments pertaining to compounds of Formula XXI, R⁴ is Hor CH₃ and R⁵ is an optionally substituted five-, six- or seven-memberedcarbocyclic or heterocyclic ring (e.g., optionally substituted phenylring). In some embodiments pertaining to compounds of Formula XXI, R⁴ isH or CH₃ and R⁵ is a phenyl ring substituted with one or more halogen(e.g., F, Cl), fluoroalkyl (e.g., CF₃), or acetylene substituents, whichsubstituents sometimes are at the 3-position, 4-position or 5-position,or a combination thereof (e.g., the 3- and 5-positions). R⁵ in certainembodiments is a C₁₋₃ alkyl substituted with an optionally substitutedphenyl, pyridyl, morpholino, pyrrolyl, piperidinyl or pyrrolidinylsubstituent, or a C₁₋₃ alkyl substituted with a hydroxyl substituent orsubstituted with a substituent —NR⁴R⁴, where R⁴ is as defined above(e.g., —N(CH₃)₂). In other embodiments, R⁵ is a C₁₋₃ alkyl substitutedwith SO₂NR₂, NRSO₂R, NRCONR₇, NRCSNR₂, NRC(═NR)NR₂, NRCOOR, NRCOR, orCONR₂, wherein each R is independently H or C1-C8 alkyl, C2-C8heteroalkyl, C2-C8 alkenyl, C2-C8 heteroalkenyl, C2-C8 alkynyl, C2-C8heteroalkynyl, C1-C8 acyl, C2-C8 heteroacyl, C6-C10 aryl, C5-C10heteroaryl, C7-C12 arylalkyl, or C6-C12 heteroarylalkyl. An R⁶substituent in certain embodiments, such as R^(6A) or R^(6B), sometimesis a halo, or a —NR⁴R⁵ substituent, such as a —NH—(C1-C6 alkyl) moiety(e.g., —NH—CH₃), for example.

Also provided herein are compounds of Formulae XXV, XXVI and XXVII:

and pharmaceutically acceptable salts, esters, prodrugs and tautomersthereof; wherein:

each Z¹, Z², Z³, and Z⁴ is N or CR³;

each of Z⁵, Z⁶, Z⁷ and Z⁸ is CR⁶;

each R³ and each R⁶ is independently H or an optionally substitutedC1-C8 alkyl, C2-C8 heteroalkyl, C2-C8 alkenyl, C2-C8 heteroalkenyl,C2-C8 alkynyl, C2-C8 heteroalkynyl, C1-C8 acyl, C2-C8 heteroacyl, C6-C10aryl, C5-C12 heteroaryl, C7-C12 arylalkyl, or C6-C12 heteroarylalkylgroup,

or each R³ and each R⁶ can be halo, OR, NR₂, NROR, NRNR₂, SR, SOR, SO₂R,SO₂NR₂, NRSO₂R, NRCONR₂, NRCSNR₂, NRC(═NR)NR₂, NRCOOR, NRCOR, CN, COOR,CONR₂, OOCR, COR, or NO₂,

wherein each R is independently H or C1-C8 alkyl, C2-C8 heteroalkyl,C2-C8 alkenyl, C2-C8 heteroalkenyl, C2-C8 alkynyl, C2-C8 heteroalkynyl,C1-C8 acyl, C2-C8 heteroacyl, C6-C10 aryl, C5-C10 heteroaryl, C7-C12arylalkyl, or C6-C12 heteroarylalkyl,

and wherein two R on the same atom or on adjacent atoms can be linked toform a 3-8 membered ring, optionally containing one or more N, O or S;

and each R group, and each ring formed by linking two R groups together,is optionally substituted with one or more substituents selected fromhalo, ═O, ═N—CN, ═N—OR′, ═NR′, OR′, NR′₂, SR′, SO₂R′, SO₂NR′₂, NR′SO₂R′,NR′CONR′₂, NR′CSNR′₂, NR′C(═NR′)NR′₂, NR′COOR′, NR′COR′, CN, COOR′,CONR′₂, OOCR′, COR′, and NO₂,

wherein each R′ is independently H, C1-C6 alkyl, C2-C6 heteroalkyl,C1-C6 acyl, C2-C6 heteroacyl, C6-C10 aryl, C5-C10 heteroaryl, C7-12arylalkyl, or C6-12 heteroarylalkyl, each of which is optionallysubstituted with one or more groups selected from halo, C1-C4 alkyl,C1-C4 heteroalkyl, C1-C6 acyl, C1-C6 heteroacyl, hydroxy, amino, and ═O;

and wherein two R′ can be linked to form a 3-7 membered ring optionallycontaining up to three heteroatoms selected from N, O and S,

R⁴ is H or optionally substituted member selected from the groupconsisting of C₁-C₆ alkyl, C2-C6 heteroalkyl, and C1-C6 acyl;

each R⁵ is independently H or an optionally substituted member selectedfrom the group consisting of C₁₋₁₀ alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀heteroalkyl, C₃₋₈ carbocyclic ring, and C₃₋₈ heterocyclic ringoptionally fused to an additional optionally substituted carbocyclic orheterocyclic; or R⁵ is a C₁₋₁₀ alkyl, C₂₋₁₀ alkenyl, or C₂₋₁₀heteroalkyl substituted with an optionally substituted C₃₋₈ carbocyclicring or C₃₋₈ heterocyclic ring; and

in each —NR⁴R⁵, R⁴ and R⁵ together with N may form an optionallysubstituted 3-8 membered ring, which may optionally contain anadditional heteroatom selected from N, O and S as a ring member;

provided that when —NR⁴R⁵ is —NHΦ, where Φ is optionally substitutedphenyl:

at least one R³ present must be a polar substituent, or if each R³ is H,then Φ must be substituted.

In some embodiments pertaining to compounds of Formulae XXV, XXVI, orXXVII, each of Z¹, Z², Z³, and Z⁴ is CR³, and at least one R³ is H, orat least two R³ are H. Often, at least one of R⁶ is H, and sometimeseach of R⁶ is H. In certain embodiments, each R³ and/or each of R⁶present is H, except that at least one R³ present is a polarsubstituent. In some embodiments, each R^(3A), R^(3C), R^(3D), and R⁶ isH and R^(3B) is a polar substituent (e.g., carboxylate bioisostere,carboxylic acid, carboxamide, or tetrazole). Embodiments described withrespect to compounds of Formulae I, II, III and IV also may be appliedto compounds of Formulae XXV, XXVI, or XXVII.

In certain embodiments pertaining to compounds of Formulae XXV, XXVI, orXXVII, R⁴ is H or CH₃ and R⁵ is an optionally substituted five-, six- orseven-membered carbocyclic or heterocyclic ring (e.g., optionallysubstituted phenyl ring). In some embodiments, R⁴ is H or CH₃ and R⁵ isa phenyl ring substituted with one or more halogen (e.g., F, Cl),fluoroalkyl (e.g., CF₃), or acetylene substituents, which substituentssometimes are at the 3-position, 4-position or 5-position, or acombination thereof (e.g., the 3- and 5-positions). R⁵ in certainembodiments is a C₁₋₃ alkyl substituted with an optionally substitutedphenyl, pyridyl, morpholino, pyrrolyl, piperidinyl or pyrrolidinylsubstituent, or a C₁₋₃ alkyl substituted with a hydroxyl substituent orsubstituted with a substituent —NR⁴R⁴, where R⁴ is as defined above(e.g., —N(CH₃)₂). In other embodiments, R⁵ is a C₁₋₃ alkyl substitutedwith SO₂NR₂, NRSO₂R, NRCONR₂, NRCSNR₂, NRC(═NR)NR₂, NRCOOR, NRCOR, orCONR₂, wherein each R is independently H or C1-C8 alkyl, C2-C8heteroalkyl, C2-C8 alkenyl, C2-C8 heteroalkenyl, C2-C8 alkynyl, C2-C8heteroalkynyl, C1-C8 acyl, C2-C8 heteroacyl, C6-C10 aryl, C5-C10heteroaryl, C7-C12 arylalkyl, or C6-C12 heteroaryl alkyl. An R⁶substituent in certain embodiments sometimes is a halo, or a —NR⁴R⁵substituent, such as a —NH—(C1-C6 alkyl) moiety (e.g., —NH—CH₃), forexample.

In some embodiments of compounds of Formula I, the invention providescompounds having activity on Pim kinases, particularly Pim1 and/or Pim2kinase. Compounds of Formula IA (and IB and IC) are inhibitors of atleast one of these Pim kinases, and are accordingly useful to treatconditions characterized by or associated with excessive Pim activity.This aspect of the invention provides compounds having the Formula IA,IB and IC, pharmaceutical compositions comprising at least one suchcompound admixed with one or more pharmaceutically acceptable excipientsand/or carriers, and methods of using these compounds to treatconditions such as the cancers described herein, as well as pain andinflammation. The compounds have this formula:

or a pharmaceutically acceptable salt thereof.

In certain preferred embodiments, the compounds of Formula IA includecompounds of Formula IB or IC:

or a pharmaceutically acceptable salt thereof.

In compounds of Formula IA, IB, and IC:

Z⁶⁰ and Z⁷⁰ are independently N or CR⁶⁰, provided at least one of themis N;

each R³⁰ and each R⁶⁰ is independently H or an optionally substitutedC1-C8 alkyl, C2-C8 heteroalkyl, C2-C8 alkenyl, C2-C8 heteroalkenyl,C2-C8 alkynyl, C2-C8 heteroalkynyl, C1-C8 acyl, C2-C8 heteroacyl, C6-C10aryl, C5-C12 heteroaryl, C7-C12 aryl alkyl, or C6-C12 heteroaryl alkylgroup,

or each R³⁰ and each R⁶⁰ can be halo, OR, NR₂, NROR, NRNR₂, SR, SOR,SO₂R, SO₂NR₂, NRSO₂R, NRCONR₂, NRCSNR₂, NRC(═NR)NR₂, NRCOOR, NRCOR, CN,COOR, CONR₂, OOCR, COR, or NO₂,

wherein each R is independently H or C1-C8 alkyl, C2-C8 heteroalkyl,C2-C8 alkenyl, C2-C8 heteroalkenyl, C2-C8 alkynyl, C2-C8 heteroalkynyl,C1-C8 acyl, C2-C8 heteroacyl, C6-C10 aryl, C5-C10 heteroaryl, C7-C12arylalkyl, or C6-C12 heteroarylalkyl,

and wherein two R on the same atom or on adjacent atoms can be linked toform a 3-8 membered ring, optionally containing one or more N, O or S;

and each R group, and each ring formed by linking two R groups together,is optionally substituted with one or more substituents selected fromhalo, ═O, ═N—CN, ═N—OR′, ═NR′, OR′, NR′₂, SR′, SO₂R′, SO₂NR′₂, NR′SO₂R′,NR′CONR′₂, NR′CSNR′₂, NR′C(═NR′)NR′₂, NR′COOR′, NR′COR′, CN, COOR′,CONR′₂, ° OCR', COR′, and NO₂,

wherein each R′ is independently H, C1-C6 alkyl, C2-C6 heteroalkyl,C1-C6 acyl, C2-C6 heteroacyl, C6-C10 aryl, C5-C10 heteroaryl, C7-12arylalkyl, or C6-12 heteroarylalkyl, each of which is optionallysubstituted with one or more groups selected from halo, C1-C4 alkyl,C1-C4 heteroalkyl, C1-C6 acyl, C1-C6 heteroacyl, hydroxy, amino, and ═O;

and wherein two R′ can be linked to form a 3-7 membered ring optionallycontaining up to three heteroatoms selected from N, O and S,

each R⁴⁰ is H or optionally substituted member selected from the groupconsisting of C₁-C₆ alkyl, C2-C6 heteroalkyl, and C1-C6 acyl;

each R⁵⁰ is independently an optionally substituted member selected fromthe group consisting of C₁₋₁₀ alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ heteroalkyl,C₃₋₈ carbocyclic ring, and C₃₋₈ heterocyclic ring optionally fused to anadditional optionally substituted carbocyclic or heterocyclic;

or R⁵⁰ can be a C₁₋₁₀ alkyl, C₂₋₁₀ alkenyl, or C₂₋₁₀ heteroalkylsubstituted with an optionally substituted C₃₋₈ carbocyclic ring or C₃₋₈heterocyclic ring;

in each —NR⁴⁰R⁵⁰, R⁴⁰ and R⁵⁰ together with N may form an optionallysubstituted 3-8 membered ring, which may optionally contain anadditional heteroatom selected from N, O and S as a ring member;

each R^(3P) represents a polar substituent;

and each Φ independently represents an optionally substituted phenyl.

Pharmaceutically acceptable salts and tautomers of the compounds ofFormulae IA, IB and IC are also included within the scope of theinvention.

In some compounds of Formula IA, Z⁶⁰ can be N while Z⁷⁰ is CH, or Z⁷⁰can be N while Z⁶⁰ is CH. In some of these compounds, R⁴⁰ is H or aC1-C6 acyl group, or a C1-C6 alkyl group. In some of these compounds,R⁵⁰ is an optionally substituted phenyl group, or R⁵⁰ can be—(CH₂)_(q)—RG, where q is an integer from 0-2 and RG represents anoptionally substituted ring selected from phenyl, 2-pyridyl, 3-pyridyl,4-pyridyl, morpholine, piperizine, piperidine, pyrrolidine, andcyclopropane. A preferred embodiment of Formula IA includes compoundswherein R⁵⁰ is optionally substituted phenyl (i.e., Φ).

In some compounds of Formula IA, each R³⁰ is independently H or halo orC1-C6 alkyl. Preferably at least one R³⁰ is H in these compounds.

R^(3P) is a polar substituent, and can be any of the polar substituentsdescribed above for compounds of Formula I. In some embodiments of thecompounds of Formula IA, R^(3P) is a triazole or imidazole ring, whichcan be substituted or unsubstituted, and is preferably bonded through acarbon atom of the triazole or imidazole ring to the fused tricyclicmoiety in Formula IA. In other embodiments, R^(3P) is a carboxylic acidor a salt, an ester or a bioisostere thereof. In some embodiments, atleast one R³ is a carboxylic acid-containing substituent or acarboxylate bioisostere, or a salt or ester thereof, for example. Insome embodiments, at least one R³ is a carboxylic acid-containingsubstituent or a salt thereof. In other embodiments, R^(3P) representsan amide group of the formula —C(O)NR⁴⁰R⁵⁰, where NR⁴⁰R⁵⁰ is as definedabove. In other embodiments, R^(3P) represents an ester group —COOR⁸⁰,wherein R⁸⁰ is H or an optionally substituted C1-C6 alkyl. Embodimentsof R^(3P) described with respect to compounds of Formula I are alsouseful herein for compounds of formula IA, IB, and IC. Embodiments ofR^(3P) described with respect to compounds of Formula I, IA, IB, and ICare also useful for compounds of Formula L, L-A and L-B.

In compounds of formula IB or IC, R³⁰ is typically H or halo. R^(3P) isfrequently a 2-imidazolyl ring or a 3-triazolyl ring, each of which canbe unsubstituted or substituted. If these rings are substituted on N,they are typically substituted with C1-C6 alkyl or C1-C6 acyl, or, ifsubstituted on a carbon atom of the ring, with halo. Unsubstituted3-triazole is a preferred group for R^(3P).

In the compounds of Formula IB or IC, Φ is an optionally substitutedphenyl, which can be unsubstituted phenyl or a phenyl substituted with1-3 substituents. In some embodiments, the substituents on the phenylring are selected from halo, cyano, CF₃, —OCF₃, COOR⁴⁰, and SO₂NR⁴⁰R⁵⁰,and one or more of these substituents can be an optionally substitutedgroup selected from C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, and C2-C6alkynyl.

In some embodiments of the invention, the compound has the structure ofFormula L, L-A and L-B. This aspect of the invention provides compoundshaving the Formula L, pharmaceutical compositions comprising at leastone such compound admixed with one or more pharmaceutically acceptableexcipients and/or carriers, and methods of using these compounds totreat conditions such as cancers, inflammation or pain, as describedherein. The compounds have this formula:

or a pharmaceutically acceptable salt thereof.

In certain preferred embodiments, the compounds of Formula L includecompounds of Formula L-A or L-B:

or a pharmaceutically acceptable salt thereof.

In compounds of Formula L, L-A and L-B:

Z⁶⁰ and Z⁷⁰ are independently N or CR⁶⁰, provided at least one of themis N;

each R³⁰ and each R⁶⁰ is independently H or an optionally substitutedC1-C8 alkyl, C2-C8 heteroalkyl, C2-C8 alkenyl, C2-C8 heteroalkenyl,C2-C8 alkynyl, C2-C8 heteroalkynyl, C1-C8 acyl, C2-C8 heteroacyl, C6-C10aryl, C5-C12 heteroaryl, C7-C12 arylalkyl, or C6-C12 heteroarylalkylgroup,

or each R³⁰ and each R⁶⁰ can be halo, OR, NR₂, NROR, NRNR₂, SR, SOR,SO₂R, SO₂NR₂, NRSO₂R, NRCONR₂, NRCSNR₂, NRC(═NR)NR₂, NRCOOR, NRCOR, CN,COOR, CONR₂, OOCR, COR, or NO₂,

wherein each R is independently H or C1-C8 alkyl, C2-C8 heteroalkyl,C2-C8 alkenyl, C2-C8 heteroalkenyl, C2-C8 alkynyl, C2-C8 heteroalkynyl,C1-C8 acyl, C2-C8 heteroacyl, C6-C10 aryl, C5-C10 heteroaryl, C7-C12arylalkyl, or C6-C12 heteroarylalkyl,

and wherein two R on the same atom or on adjacent atoms can be linked toform a 3-8 membered ring, optionally containing one or more N, O or S;

and each R group, and each ring formed by linking two R groups together,is optionally substituted with one or more substituents selected fromhalo, ═O, ═N—CN, ═N—OR′, ═NR′, OR′, NR′₂, SR′, SO₂R′, SO₂NR′₂, NR′SO₂R′,NR′CONR′₂, NR′CSNR′₂, NR′C(═NR′)NR′₂, NR′COOR′, NR′COR′, CN, COOR′,CONR′₂, OOCR′, COR′, and NO₂,

wherein each R′ is independently H, C1-C6 alkyl, C2-C6 heteroalkyl,C1-C6 acyl, C2-C6 heteroacyl, C6-C10 aryl, C5-C10 heteroaryl, C7-12arylalkyl, or C6-12 heteroarylalkyl, each of which is optionallysubstituted with one or more groups selected from halo, C1-C4 alkyl,C1-C4 heteroalkyl, C1-C6 acyl, C1-C6 heteroacyl, hydroxy, amino, and ═O;

and wherein two R′ can be linked to form a 3-7 membered ring optionallycontaining up to three heteroatoms selected from N, O and S,

each R^(3P) represents a polar substituent;

and each W represents an optionally substituted aryl, heteroaryl, orC₃₋₈ cycloalkyl ring.

Pharmaceutically acceptable salts and tautomers of the compounds ofFormulae L, L-A and L-B are also included within the scope of theinvention.

In some embodiments of formula L, L-A and L-B, each R3P represents anoptionally substituted imidazole or triazole ring.

In some compounds of Formula L, Z60 can be N while Z70 is CH, or Z70 canbe N while Z60 is CH.

In some embodiments of formula L, L-A and L-B, W represents a monocyclic6-membered aromatic or 5-6 membered heteroaromatic ring, or a fusedbicyclic 8-10 membered aromatic or heteroaromatic ring, each of whichmay be optionally substituted. In some such embodiments, W represents anoptionally substituted aromatic or heteraromatic ring selected from thegroup consisting of phenyl, naphthyl, pyridine, pyrimidine, pyridazine,thiophene, oxazole, isoxazole, imidazole, pyrazole, pyrrole, thiazole,and isothiazole.

A preferred embodiment of Formula L includes compounds wherein W isoptionally substituted phenyl ring. In other embodiments, W representsan optionally substituted C3-8 cycloalkyl ring; sometimes W iscyclopropyl.

In some embodiments of Formula L, each R30 is independently H, halo orC1-C6 alkyl. Preferably at least one R30 is H in these compounds.

R3P is a polar substituent, and can be any of the polar substituentsdescribed above for compounds of Formula IA, IB and IC. In someembodiments of Formula L, R3P is a triazole or imidazole ring, which canbe substituted or unsubstituted, and is preferably bonded through acarbon atom of the triazole or imidazole ring to the fused tricyclicmoiety in Formula L.

In compounds of formula L-A or L-B, R30 is typically H or halo. R3P isfrequently a 2-imidazolyl ring or a 3-triazolyl ring, each of which canbe unsubstituted or substituted. If these rings are substituted on N,they are typically substituted with C1-C6 alkyl or C1-C6 acyl, or, ifsubstituted on a carbon atom of the ring, with halo. Unsubstituted3-triazole is a preferred group for R3P.

In some embodiments of Formula L-A or L-B, W is an optionallysubstituted phenyl, which can be unsubstituted phenyl or a phenylsubstituted with 1-3 substituents. In some embodiments, the substituentson the phenyl ring are selected from halo, cyano, CF3, —OCF3, COOR40,and SO2NR40R50, and one or more of these substituents can be anoptionally substituted group selected from C1-C6 alkyl, C1-C6 alkoxy,C2-C6 alkenyl, and C2-C6 alkynyl, wherein each of R40 and R50 aredefined as for formula IA.

Also provided herein is a pharmaceutical composition comprising acompound of any of the Formulae described herein, including Formula IA,IB, IC, L, L-A and L-B, and at least one pharmaceutically acceptablecarrier or excipient, or two or more pharmaceutically acceptablecarriers and/or excipients. It is understood that the compounds ofFormula I can include compounds of Formula IA, IB and IC. Pharmaceuticalcompositions can be utilized in treatments described herein.

Provided also are methods for identifying a candidate molecule thatinteracts with a CK2, Pim or Flt protein, which comprise: contacting acomposition containing a CK2, Pim or Flt protein kinase and a compounddescribed herein with a candidate molecule under conditions in which thecompound and the protein kinase interact, and determining whether theamount of the compound that interacts with the protein kinase ismodulated relative to a control interaction between the compound and theprotein kinase without the candidate molecule, whereby a candidatemolecule that modulates the amount of the compound interacting with theprotein kinase relative to the control interaction is identified as acandidate molecule that interacts with the protein kinase.

In certain embodiments the protein is in a cell or in a cell-freesystem. The protein, the compound or the molecule in some embodiments isin association with a solid phase. In certain embodiments, theinteraction between the compound and the protein is detected via adetectable label, where in some embodiments the protein comprises adetectable label and in certain embodiments the compound comprises adetectable label. The interaction between the compound and the proteinsometimes is detected without a detectable label.

In certain embodiments, the protein is a CK2 protein, such as a CK2protein comprising the amino acid sequence of SEQ ID NO: 1, 2 or 3 or asubstantially identical variant thereof, for example.

SEQ ID NO: 1(NP_001886; casein kinase II alpha 1 subunit isoform a [Homo sapiens])  1 msgpvpsrar vytdvnthrp reywdyeshv vewgnqddyq lvrklgrgky sevfeainit 61 nnekvvvkil kpvkkkkikr eikilenlrg gpniitladi vkdpvsrtpa lvfehvnntd121 fkqlyqtltd ydirfymyei lkaldychsm gimhrdvkph nvmidhehrk lrlidwglae181 fyhpgqeynv rvasryfkgp ellvdyqmyd ysldmwslgc mlasmifrke pffhghdnyd241 qlvriakvlg tedlydyidk ynieldprfn dilgrhsrkr werfvhsenq hlvspealdf301 ldkllrydhq srltareame hpyfytvvkd qarmgsssmp ggstpvssan mmsgissvpt361 psplgplags pviaaanplg mpvpaaagaq q SEQ ID NO: 2(NP_808227; casein kinase II alpha 1 subunit isoform a [Homo sapiens])  1 msgpvpsrar vytdvnthrp reywdyeshv vewgnqddyq lvrklgrgky sevfeainit 61 nnekvvvkil kpvkkkkikr eikilenlrg gpniitladi vkdpvsrtpa lvfehvnntd121 fkqlyqtltd ydirfymyei lkaldychsm gimhrdvkph nvmidhehrk lrlidwglae181 fyhpgqeynv rvasryfkgp ellvdyqmyd ysldmwslgc mlasmifrke pffhghdnyd241 qlvriakvlg tedlydyidk ynieldprfn dilgrhsrkr werfvhsenq hlvspealdf301 ldkllrydhq srltareame hpyfytvvkd qarmgsssmp ggstpvssan mmsgissvpt361 psplgplags pviaaanplg mpvpaaagaq q SEQ ID NO: 3(NP_808228; casein kinase II alpha 1 subunit isoform b [Homo sapiens])  1 myeilkaldy chsmgimhrd vkphnvmidh ehrklrlidw glaefyhpgq eynvrvasry 61 fkgpellvdy qmydysldmw slgcmlasmi frkepffhgh dnydqlvria kvlgtedlyd121 yidkynield prfndilgrh srkrwerfvh senqhlvspe aldfldkllr ydhqsrltar181 eamehpyfyt vvkdqarmgs ssmpggstpv ssanmmsgis svptpsplgp lagspviaaa241 nplgmpvpaa agaqq

Also provided are methods for modulating the activity of a CK2 protein,Pim protein, or Flt protein which comprise contacting a systemcomprising the protein with a compound described herein in an amounteffective for modulating the activity of the protein. In certainembodiments the activity of the protein is inhibited, and sometimes theprotein is a CK2 protein, such as a CK2 protein comprising the aminoacid sequence of SEQ ID NO: 1, 2 or 3 or a substantially identicalvariant thereof, for example. In other embodiments the protein is a Pimprotein or a Flt protein. In certain embodiments, the system is a cell,and in other embodiments the system is a cell-free system. The proteinor the compound may be in association with a solid phase in certainembodiments.

Provided also are methods for inhibiting cell proliferation, whichcomprise contacting cells with a compound described herein in an amounteffective to inhibit proliferation of the cells. The cells sometimes arein a cell line, such as a cancer cell line (e.g., breast cancer,prostate cancer, pancreatic cancer, lung cancer, hemopoietic cancer,colorectal cancer, skin cancer, ovary cancer cell line), for example. Insome embodiments, the cancer cell line is a breast cancer, prostatecancer or pancreatic cancer cell line. The cells sometimes are in atissue, can be in a subject, at times are in a tumor, and sometimes arein a tumor in a subject. In certain embodiments, the method furthercomprises inducing cell apoptosis. Cells sometimes are from a subjecthaving macular degeneration.

Also provided are methods for treating a condition related to aberrantcell proliferation, which comprise administering a compound describedherein to a subject in need thereof in an amount effective to treat thecell proliferative condition. In certain embodiments the cellproliferative condition is a tumor-associated cancer. The cancersometimes is of the breast, prostate, pancreas, lung, colorectum, skin,or ovary. In some embodiments, the cell proliferative condition is anon-tumor cancer, such as a hematopoietic cancer, for example. In otherembodiments, the cell proliferative condition is macular degeneration insome embodiments.

Provided also are methods for treating cancer or an inflammatorydisorder in a subject in need of such treatment, comprising:administering to the subject a therapeutically effective amount of atherapeutic agent useful for treating such disorder; and administeringto the subject a molecule that inhibits CK2, Pim or Flt in an amountthat is effective to enhance a desired effect of the therapeutic agent.In certain embodiments, the molecule that inhibits CK2, Pim or Flt is acompound of Formula I, IA, IB, IC, L, L-A or L-B as described herein, ora pharmaceutically acceptable salt thereof. In some embodiments, themolecule that inhibits CK2, Pim or Flt is a known compound shown above,or a compound in one of the Tables provided herein, or apharmaceutically acceptable salt of one of these compounds. In someembodiments, the desired effect of the therapeutic agent that isenhanced by the molecule that inhibits CK2, Pim or Flt is a reduction incell proliferation. In certain embodiments, the desired effect of thetherapeutic agent that is enhanced by the molecule that inhibits CK2,Pim or Flt is an increase in apoptosis in at least one type of cell.

In some embodiments, the therapeutic agent and the molecule thatinhibits CK2, Pim or Flt are administered at substantially the sametime. The therapeutic agent and molecule that inhibits CK2, Pim or Fltsometimes are used concurrently by the subject. The therapeutic agentand the molecule that inhibits CK2, Pim or Flt are combined into onepharmaceutical composition in certain embodiments.

Also provided are compositions of matter comprising a compound describedherein and an isolated protein. The protein sometimes is a CK2 protein,such as a CK2 protein comprising the amino acid sequence of SEQ ID NO:1, 2 or 3 or a substantially identical variant thereof, for example. Insome embodiments, the protein is a Pim protein. In other embodiments,the protein is a Flt protein. Certain compositions comprise a compounddescribed herein in combination with a cell. The cell may be from a cellline, such as a cancer cell line. In the latter embodiments, the cancercell line is sometimes a breast cancer, prostate cancer, pancreaticcancer, lung cancer, hemopoietic cancer, colorectal cancer, skin cancer,ovary cancer cell line.

These and other embodiments of the invention are described in thedescription that follows.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 depicts assay data showing inhibition of CK2 activity.

FIGS. 2A and 2B show mean plasma concentrations of compounds describedherein over time after intravenous and oral administration to ICR mice.

FIGS. 3A and 3B show tumor volume over time and body weight over time,respectively, in tumor-bearing xenograft animals administered a compounddescribed herein. FIGS. 3C and 3D illustrate effects of the compound ontumors in individual animals.

FIGS. 4A and 4B show tumor volume over time and body weight over time,respectively, in tumor-bearing xenograft animals administered a compounddescribed herein.

MODES OF CARRYING OUT THE INVENTION

Compounds of the Formulae provided herein, including compounds ofFormulae IA, IB, IC, L, L-A or L-B, can exert biological activities thatinclude, but are not limited to, inhibiting cell proliferation.Compounds of such Formulae can modulate CK2 activity, Pim activityand/or Flt activity, for example. Such compounds therefore can beutilized in multiple applications by a person of ordinary skill in theart. For example, compounds described herein may find uses that include,but are not limited to, (i) modulation of protein kinase activity (e.g.,CK2 activity), (ii) modulation of Pim activity (e.g., PIM-1 activity),(iii) modulation of FMS-like tyrosine kinase (Flt) activity (e.g., Flt-3activity), (iv) modulation of cell proliferation, (v) modulation ofapoptosis, and (vi) treatments of cell proliferation related disorders,pain or inflammation (e.g., administration alone or co-administrationwith another molecule).

“Optionally substituted” as used herein indicates that the particulargroup or groups being described may have no non-hydrogen substituents,or the group or groups may have one or more non-hydrogen substituents.If not otherwise specified, the total number of such substituents thatmay be present is equal to the number of H atoms present on theunsubstituted form of the group being described. Where an optionalsubstituent is attached via a double bond, such as a carbonyl oxygen(═O), the group takes up two available valences, so the total number ofsubstituents that may be included is reduced according to the number ofavailable valences.

The compounds of the invention often have ionizable groups so as to becapable of preparation as salts. In that case, wherever reference ismade to the compound, it is understood in the art that apharmaceutically acceptable salt may also be used. These salts may beacid addition salts involving inorganic or organic acids or the saltsmay, in the case of acidic forms of the compounds of the invention beprepared from inorganic or organic bases. Frequently, the compounds areprepared or used as pharmaceutically acceptable salts prepared asaddition products of pharmaceutically acceptable acids or bases.Suitable pharmaceutically acceptable acids and bases are well-known inthe art, such as hydrochloric, sulphuric, hydrobromic, acetic, lactic,citric, or tartaric acids for forming acid addition salts, and potassiumhydroxide, sodium hydroxide, ammonium hydroxide, caffeine, variousamines, and the like for forming basic salts. Methods for preparation ofthe appropriate salts are well-established in the art. In some cases,the compounds may contain both an acidic and a basic functional group,in which case they may have two ionized groups and yet have no netcharge.

In some cases, the compounds of the invention contain one or more chiralcenters. The invention includes each of the isolated stereoisomericforms as well as mixtures of stereoisomers in varying degrees of chiralpurity, including racemic mixtures. It also encompasses the variousdiastereomers and tautomers that can be formed. The compounds of theinvention may also exist in more than one tautomeric form; the depictionherein of one tautomer is for convenience only, and is also understoodto encompass other tautomers of the form shown.

As used herein, the terms “alkyl,” “alkenyl” and “alkynyl” includestraight-chain, branched-chain and cyclic monovalent hydrocarbylradicals, and combinations of these, which contain only C and H whenthey are unsubstituted. Examples include methyl, ethyl, isobutyl,cyclohexyl, cyclopentylethyl, 2-propenyl, 3-butynyl, and the like. Thetotal number of carbon atoms in each such group is sometimes describedherein, e.g., when the group can contain up to ten carbon atoms it canbe represented as 1-10C or as C1-C10 or C1-10. When heteroatoms (N, Oand S typically) are allowed to replace carbon atoms as in heteroalkylgroups, for example, the numbers describing the group, though stillwritten as e.g. C₁-C₆, represent the sum of the number of carbon atomsin the group plus the number of such heteroatoms that are included asreplacements for carbon atoms in the backbone of the ring or chain beingdescribed.

Typically, the alkyl, alkenyl and alkynyl substituents of the inventioncontain 1-10C (alkyl) or 2-10C (alkenyl or alkynyl). Preferably theycontain 1-8C (alkyl) or 2-8C (alkenyl or alkynyl). Sometimes theycontain 1-4C (alkyl) or 2-4C (alkenyl or alkynyl). A single group caninclude more than one type of multiple bond, or more than one multiplebond; such groups are included within the definition of the term“alkenyl” when they contain at least one carbon-carbon double bond, andare included within the term “alkynyl” when they contain at least onecarbon-carbon triple bond.

Alkyl, alkenyl and alkynyl groups are often optionally substituted tothe extent that such substitution makes sense chemically. Typicalsubstituents include, but are not limited to, halo, ═O, ═N—CN, ═NR, OR,NR₂, SR, SO₂R, SO₂NR₂, NRSO₂R, NRCONR₂, NRCSNR₂, NRC(═NR)NR₂, NRCOOR,NRCOR, CN, C≡CR, COOR, CONR₂, OOCR, COR, and NO₂, wherein each R isindependently H, C1-C8 alkyl, C2-C8 heteroalkyl, C1-C8 acyl, C2-C8heteroacyl, C2-C8 alkenyl, C2-C8 heteroalkenyl, C2-C8 alkynyl, C2-C8heteroalkynyl, C6-C10 aryl, or C5-C10 heteroaryl, and each R isoptionally substituted with halo, ═O, ═N—CN, ═N—OR′, ═NR′, OR′, NR′₂,SR′, SO₂R′, SO₂NR′₂, NR′SO₂R′, NR′CONR′₂, NR′CSNR′₂, NR′C(═NR′)NR′₂,NR′COOR′, NR′COR′, CN, C≡CR′, COOR′, CONR′₂, OOCR′, COR′, and NO₂,wherein each R′ is independently H, C1-C8 alkyl, C2-C8 heteroalkyl,C1-C8 acyl, C2-C8 heteroacyl, C6-C10 aryl or C5-C10 heteroaryl. Alkyl,alkenyl and alkynyl groups can also be substituted by C1-C8 acyl, C2-C8heteroacyl, C6-C10 aryl or C5-C10 heteroaryl, each of which can besubstituted by the substituents that are appropriate for the particulargroup.

“Acetylene” substituents are 2-10C alkynyl groups that are optionallysubstituted, and are of the formula —C≡C—R^(a), wherein R^(a) is H orC1-C8 alkyl, C2-C8 heteroalkyl, C2-C8 alkenyl, C2-C8 heteroalkenyl,C2-C8 alkynyl, C2-C8 heteroalkynyl, C1-C8 acyl, C2-C8 heteroacyl, C6-C10aryl, C5-C10 heteroaryl, C7-C12 arylalkyl, or C6-C12 heteroarylalkyl,

and each Ra group is optionally substituted with one or moresubstituents selected from halo, ═O, ═N—CN, ═N—OR′, ═NR′, OR′, NR′2,SR′, SO2R′, SO2NR′2, NR′SO2R′, NR′CONR′2, NR′CSNR′2, NR′C(═NR′)NR′2,NR′COOR′, NR′COR′, CN, COOR′, CONR′2, OOCR′, COR′, and NO2, wherein eachR′ is independently H, C1-C6 alkyl, C2-C6 heteroalkyl, C1-C6 acyl, C2-C6heteroacyl, C6-C10 aryl, C5-C10 heteroaryl, C7-12 arylalkyl, or C6-12heteroarylalkyl, each of which is optionally substituted with one ormore groups selected from halo, C1-C4 alkyl, C1-C4 heteroalkyl, C1-C6acyl, C1-C6 heteroacyl, hydroxy, amino, and ═O; and wherein two R′ canbe linked to form a 3-7 membered ring optionally containing up to threeheteroatoms selected from N, O and S. In some embodiments, R^(a) of—C≡C—R^(a) is H or Me.

“Heteroalkyl”, “heteroalkenyl”, and “heteroalkynyl” and the like aredefined similarly to the corresponding hydrocarbyl (alkyl, alkenyl andalkynyl) groups, but the ‘hetero’ terms refer to groups that contain 1-3O, S or N heteroatoms or combinations thereof within the backboneresidue; thus at least one carbon atom of a corresponding alkyl,alkenyl, or alkynyl group is replaced by one of the specifiedheteroatoms to form a heteroalkyl, heteroalkenyl, or heteroalkynylgroup. The typical and preferred sizes for heteroforms of alkyl, alkenyland alkynyl groups are generally the same as for the correspondinghydrocarbyl groups, and the substituents that may be present on theheteroforms are the same as those described above for the hydrocarbylgroups. For reasons of chemical stability, it is also understood that,unless otherwise specified, such groups do not include more than twocontiguous heteroatoms except where an oxo group is present on N or S asin a nitro or sulfonyl group.

While “alkyl” as used herein includes cycloalkyl and cycloalkylalkylgroups, the term “cycloalkyl” may be used herein to describe acarbocyclic non-aromatic group that is connected via a ring carbon atom,and “cycloalkylalkyl” may be used to describe a carbocyclic non-aromaticgroup that is connected to the molecule through an alkyl linker.Similarly, “heterocyclyl” may be used to describe a non-aromatic cyclicgroup that contains at least one heteroatom as a ring member and that isconnected to the molecule via a ring atom, which may be C or N; and“heterocyclylalkyl” may be used to describe such a group that isconnected to another molecule through a linker. The sizes andsubstituents that are suitable for the cycloalkyl, cycloalkylalkyl,heterocyclyl, and heterocyclylalkyl groups are the same as thosedescribed above for alkyl groups. As used herein, these terms alsoinclude rings that contain a double bond or two, as long as the ring isnot aromatic.

As used herein, “acyl” encompasses groups comprising an alkyl, alkenyl,alkynyl, aryl or arylalkyl radical attached at one of the two availablevalence positions of a carbonyl carbon atom, and heteroacyl refers tothe corresponding groups wherein at least one carbon other than thecarbonyl carbon has been replaced by a heteroatom chosen from N, O andS. Thus heteroacyl includes, for example, —C(═O)OR and —C(═O)NR₂ as wellas —C(═O)-heteroaryl.

Acyl and heteroacyl groups are bonded to any group or molecule to whichthey are attached through the open valence of the carbonyl carbon atom.Typically, they are C1-C8 acyl groups, which include formyl, acetyl,pivaloyl, and benzoyl, and C2-C8 heteroacyl groups, which includemethoxyacetyl, ethoxycarbonyl, and 4-pyridinoyl. The hydrocarbyl groups,aryl groups, and heteroforms of such groups that comprise an acyl orheteroacyl group can be substituted with the substituents describedherein as generally suitable substituents for each of the correspondingcomponent of the acyl or heteroacyl group.

“Aromatic” moiety or “aryl” moiety refers to a monocyclic or fusedbicyclic moiety having the well-known characteristics of aromaticity;examples include phenyl and naphthyl. Similarly, “heteroaromatic” and“heteroaryl” refer to such monocyclic or fused bicyclic ring systemswhich contain as ring members one or more heteroatoms selected from O, Sand N. The inclusion of a heteroatom permits aromaticity in 5-memberedrings as well as 6-membered rings. Typical heteroaromatic systemsinclude monocyclic C5-C6 aromatic groups such as pyridyl, pyrimidyl,pyrazinyl, thienyl, furanyl, pyrrolyl, pyrazolyl, thiazolyl, oxazolyl,and imidazolyl and the fused bicyclic moieties formed by fusing one ofthese monocyclic groups with a phenyl ring or with any of theheteroaromatic monocyclic groups to form a C8-C10 bicyclic group such asindolyl, benzimidazolyl, indazolyl, benzotriazolyl, isoquinolyl,quinolyl, benzothiazolyl, benzofuranyl, pyrazolopyridyl, quinazolinyl,quinoxalinyl, cinnolinyl, and the like. Any monocyclic or fused ringbicyclic system which has the characteristics of aromaticity in terms ofelectron distribution throughout the ring system is included in thisdefinition. It also includes bicyclic groups where at least the ringwhich is directly attached to the remainder of the molecule has thecharacteristics of aromaticity. Typically, the ring systems contain 5-12ring member atoms. Preferably the monocyclic aryls contain 6 ringmembers and monocylic heteroaryls contain 5-6 ring members, and thebicyclic aryls and heteroaryls contain 8-10 ring members.

Aryl and heteroaryl moieties may be substituted with a variety ofsubstituents including C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, C5-C12aryl, C1-C8 acyl, and heteroforms of these, each of which can itself befurther substituted; other substituents for aryl and heteroaryl moietiesinclude halo, OR, NR₂, SR, SO₂R, SO₂NR₂, NRSO₂R, NRCONR₂, NRCSNR₂,NRC(═NR)NR₂, NRCOOR, NRCOR, CN, C≡CR, COOR, CONR₂, OOCR, COR, and NO₂,wherein each R is independently H, C1-C8 alkyl, C2-C8 heteroalkyl, C2-C8alkenyl, C2-C8 heteroalkenyl, C2-C8 alkynyl, C2-C8 heteroalkynyl, C6-C10aryl, C5-C10 heteroaryl, C7-C12 arylalkyl, or C6-C12 heteroarylalkyl,and each R is optionally substituted as described above for alkylgroups. The substituent groups on an aryl or heteroaryl group may ofcourse be further substituted with the groups described herein assuitable for each type of such substituents or for each component of thesubstituent. Thus, for example, an arylalkyl substituent may besubstituted on the aryl portion with substituents described herein astypical for aryl groups, and it may be further substituted on the alkylportion with substituents described herein as typical or suitable foralkyl groups.

Similarly, “arylalkyl” and “heteroarylalkyl” refer to aromatic andheteroaromatic ring systems which are bonded to their attachment pointthrough a linking group such as an alkylene, including substituted orunsubstituted, saturated or unsaturated, cyclic or acyclic linkers.Typically the linker is C1-C8 alkyl or a hetero form thereof. Theselinkers may also include a carbonyl group, thus making them able toprovide substituents as an acyl or heteroacyl moiety. An aryl orheteroaryl ring in an arylalkyl or heteroarylalkyl group may besubstituted with the same substituents described above for aryl groups.Preferably, an arylalkyl group includes a phenyl ring optionallysubstituted with the groups defined above for aryl groups and a C1-C4alkylene that is unsubstituted or is substituted with one or two C1-C4alkyl groups or heteroalkyl groups, where the alkyl or heteroalkylgroups can optionally cyclize to form a ring such as cyclopropane,dioxolane, or oxacyclopentane. Similarly, a heteroarylalkyl grouppreferably includes a C5-C6 monocyclic heteroaryl group that isoptionally substituted with the groups described above as substituentstypical on aryl groups and a C1-C4 alkylene that is unsubstituted or issubstituted with one or two C1-C4 alkyl groups or heteroalkyl groups, orit includes an optionally substituted phenyl ring or C5-C6 monocyclicheteroaryl and a C1-C4 heteroalkylene that is unsubstituted or issubstituted with one or two C1-C4 alkyl or heteroalkyl groups, where thealkyl or heteroalkyl groups can optionally cyclize to form a ring suchas cyclopropane, dioxolane, or oxacyclopentane.

Where an arylalkyl or heteroarylalkyl group is described as optionallysubstituted, the substituents may be on either the alkyl or heteroalkylportion or on the aryl or heteroaryl portion of the group. Thesubstituents optionally present on the alkyl or heteroalkyl portion arethe same as those described above for alkyl groups generally; thesubstituents optionally present on the aryl or heteroaryl portion arethe same as those described above for aryl groups generally.

“Arylalkyl” groups as used herein are hydrocarbyl groups if they areunsubstituted, and are described by the total number of carbon atoms inthe ring and alkylene or similar linker. Thus a benzyl group is aC7-arylalkyl group, and phenylethyl is a C8-arylalkyl.

“Heteroarylalkyl” as described above refers to a moiety comprising anaryl group that is attached through a linking group, and differs from“arylalkyl” in that at least one ring atom of the aryl moiety or oneatom in the linking group is a heteroatom selected from N, O and S. Theheteroarylalkyl groups are described herein according to the totalnumber of atoms in the ring and linker combined, and they include arylgroups linked through a heteroalkyl linker; heteroaryl groups linkedthrough a hydrocarbyl linker such as an alkylene; and heteroaryl groupslinked through a heteroalkyl linker Thus, for example,C7-heteroarylalkyl would include pyridylmethyl, phenoxy, andN-pyrrolylmethoxy.

“Alkylene” as used herein refers to a divalent hydrocarbyl group;because it is divalent, it can link two other groups together. Typicallyit refers to —(CH₂)_(N)— where n is 1-8 and preferably n is 1-4, thoughwhere specified, an alkylene can also be substituted by other groups,and can be of other lengths, and the open valences need not be atopposite ends of a chain. Thus —CH(Me)- and —C(Me)₂- may also bereferred to as alkylenes, as can a cyclic group such ascyclopropan-1,1-diyl. Where an alkylene group is substituted, thesubstituents include those typically present on alkyl groups asdescribed herein.

In general, any alkyl, alkenyl, alkynyl, acyl, or aryl or arylalkylgroup or any heteroform of one of these groups that is contained in asubstituent may itself optionally be substituted by additionalsubstituents. The nature of these substituents is similar to thoserecited with regard to the primary substituents themselves if thesubstituents are not otherwise described. Thus, where an embodiment of,for example, R⁷ is alkyl, this alkyl may optionally be substituted bythe remaining substituents listed as embodiments for R⁷ where this makeschemical sense, and where this does not undermine the size limitprovided for the alkyl per se; e.g., alkyl substituted by alkyl or byalkenyl would simply extend the upper limit of carbon atoms for theseembodiments, and is not included. However, alkyl substituted by aryl,amino, alkoxy, ═O, and the like would be included within the scope ofthe invention, and the atoms of these substituent groups are not countedin the number used to describe the alkyl, alkenyl, etc. group that isbeing described. Where no number of substituents is specified, each suchalkyl, alkenyl, alkynyl, acyl, or aryl group may be substituted with anumber of substituents according to its available valences; inparticular, any of these groups may be substituted with fluorine atomsat any or all of its available valences, for example.

“Heteroform” as used herein refers to a derivative of a group such as analkyl, aryl, or acyl, wherein at least one carbon atom of the designatedcarbocyclic group has been replaced by a heteroatom selected from N, Oand S. Thus the heteroforms of alkyl, alkenyl, alkynyl, acyl, aryl, andarylalkyl are heteroalkyl, heteroalkenyl, heteroalkynyl, heteroacyl,heteroaryl, and heteroarylalkyl, respectively. It is understood that nomore than two N, O or S atoms are ordinarily connected sequentially,except where an oxo group is attached to N or S to form a nitro orsulfonyl group.

“Halo”, as used herein includes fluoro, chloro, bromo and iodo. Fluoroand chloro are often preferred.

“Amino” as used herein refers to NH₂, but where an amino is described as“substituted” or “optionally substituted”, the term includes NR′R″wherein each R′ and R″ is independently H, or is an alkyl, alkenyl,alkynyl, acyl, aryl, or arylalkyl group or a heteroform of one of thesegroups, and each of the alkyl, alkenyl, alkynyl, acyl, aryl, orarylalkyl groups or heteroforms of one of these groups is optionallysubstituted with the substituents described herein as suitable for thecorresponding group. The term also includes forms wherein R′ and R″ arelinked together to form a 3-8 membered ring which may be saturated,unsaturated or aromatic and which contains 1-3 heteroatoms independentlyselected from N, O and S as ring members, and which is optionallysubstituted with the substituents described as suitable for alkyl groupsor, if NR′R″ is an aromatic group, it is optionally substituted with thesubstituents described as typical for heteroaryl groups.

As used herein, the term “carbocycle” refers to a cyclic compoundcontaining only carbon atoms in the ring, whereas a “heterocycle” refersto a cyclic compound comprising a heteroatom. The carbocyclic andheterocyclic structures encompass compounds having monocyclic, bicyclicor multiple ring systems. Carbocyclic and heterocyclic rings may besaturated, partially unsaturated, or aromatic.

As used herein, the term “heteroatom” refers to any atom that is notcarbon or hydrogen, such as nitrogen, oxygen or sulfur.

Illustrative examples of heterocycles include but are not limited totetrahydrofuran, 1,3-dioxolane, 2,3-dihydrofuran, pyran,tetrahydropyran, benzofuran, isobenzofuran, 1,3-dihydro-isobenzofuran,isoxazole, 4,5-dihydroisoxazole, piperidine, pyrrolidine,pyrrolidin-2-one, pyrrole, pyridine, pyrimidine, octahydro-pyrrolo[3,4b]pyridine, piperazine, pyrazine, morpholine, thiomorpholine, imidazole,imidazolidine 2,4-dione, 1,3-dihydrobenzimidazol-2-one, indole,thiazole, benzothiazole, thiadiazole, thiophene, tetrahydro thiophene1,1-dioxide, diazepine, triazole, guanidine, diazabicyclo[2.2.1]heptane,2,5-diazabicyclo[2.2.1]heptane, 2,3,4,4a,9,9a-hexahydro-1H-β-carboline,oxirane, oxetane, tetrahydropyran, dioxane, lactones, aziridine,azetidine, piperidine, lactams, and may also encompass heteroaryls.Other illustrative examples of heteroaryls include but are not limitedto furan, pyrrole, pyridine, pyrimidine, imidazole, benzimidazole andtriazole.

As used herein, the term “inorganic substituent” refers to substituentsthat do not contain carbon or contain carbon bound to elements otherthan hydrogen (e.g., elemental carbon, carbon monoxide, carbon dioxide,and carbonate). Examples of inorganic substituents include but are notlimited to nitro, halogen, azido, cyano, sulfonyls, sulfinyls,sulfonates, phosphates, etc.

The terms “treat” and “treating” as used herein refer to ameliorating,alleviating, lessening, and removing symptoms of a disease or condition.A candidate molecule or compound described herein may be in atherapeutically effective amount in a formulation or medicament, whichis an amount that can lead to a biological effect, such as apoptosis ofcertain cells (e.g., cancer cells), reduction of proliferation ofcertain cells, or lead to ameliorating, alleviating, lessening, orremoving symptoms of a disease or condition, for example. The terms alsocan refer to reducing or stopping a cell proliferation rate (e.g.,slowing or halting tumor growth) or reducing the number of proliferatingcancer cells (e.g., removing part or all of a tumor). These terms alsoare applicable to reducing a titre of a microorganism in a system (i.e.,cell, tissue, or subject) infected with a microorganism, reducing therate of microbial propagation, reducing the number of symptoms or aneffect of a symptom associated with the microbial infection, and/orremoving detectable amounts of the microbe from the system. Examples ofmicroorganism include but are not limited to virus, bacterium andfungus. Thus the invention provides methods for treating protozoaldisorders such as protozoan parasitosis, including infection byparasitic protozoa responsible for neurological disorders such asschizophrenia, paranoia, and encephalitis in immunocompromised patients,as well as Chagas' disease. It also provides methods to treat variousviral diseases, including human immunodeficiency virus type 1 (HIV-1),human papilloma viruses (HPVs), herpes simplex virus (HSV), Epstein-Barrvirus (EBV), human cytomegalovirus, hepatitis C and B viruses, influenzavirus, Borna disease virus, adenovirus, coxsackievirus, coronavirus andvaricella zoster virus. The methods for treating these disorderscomprises administering to a subject in need thereof an effective amountof a CK2 inhibitor of Formula A.

“Treating” or “treatment” as used herein with respect to cancers or cellproliferative disorders also covers the treatment of a disease-state ina human, which disease-state is characterized by abnormal, excessiveand/or undesired cellular proliferation, and includes at least one of:(i) preventing the disease-state from occurring in a human, inparticular, when such human is predisposed to the disease-state but hasnot yet been diagnosed as having it; (ii) inhibiting the disease-state,i.e., arresting its development; (iii) inhibiting spread of the diseasestate to new loci, e.g., slowing or preventing metastasis of a tumor;and (iv) relieving the disease-state, i.e., causing regression of thedisease-state.

‘Treating’ or ‘treatment’ with regard to inflammatory conditionsincludes prevention of inflammation in a subject where inflammation isexpected to occur, or reduction of the extent or duration of one or moreof the symptoms of inflammation in a subject having symptoms ofinflammation such as redness, swelling, pain associated with these, orelevated temperature.

As used herein, the term “apoptosis” refers to an intrinsic cellself-destruction or suicide program. In response to a triggeringstimulus, cells undergo a cascade of events including cell shrinkage,blebbing of cell membranes and chromatic condensation and fragmentation.These events culminate in cell conversion to clusters of membrane-boundparticles (apoptotic bodies), which are thereafter engulfed bymacrophages.

The invention in part provides pharmaceutical compositions comprising atleast one compound within the scope of the invention as describedherein, and methods of using compounds described herein. For example,the invention in part provides methods for identifying a candidatemolecule that interacts with a CK2, Pim or Flt protein, which comprisescontacting a composition containing a CK2, Pim or Flt protein and amolecule described herein with a candidate molecule and determiningwhether the amount of the molecule described herein that interacts withthe protein is modulated, whereby a candidate molecule that modulatesthe amount of the molecule described herein that interacts with theprotein is identified as a candidate molecule that interacts with theprotein.

Provided also are methods for modulating a protein kinase activity.Protein kinases catalyze the transfer of a gamma phosphate fromadenosine triphosphate to a serine or threonine amino acid(serine/threonine protein kinase), tyrosine amino acid (tyrosine proteinkinase), tyrosine, serine or threonine (dual specificity protein kinase)or histidine amino acid (histidine protein kinase) in a peptide orprotein substrate. Thus, included herein are methods which comprisecontacting a system comprising a protein kinase protein with a compounddescribed herein in an amount effective for modulating (e.g.,inhibiting) the activity of the protein kinase. In some embodiments, theactivity of the protein kinase is the catalytic activity of the protein(e.g., catalyzing the transfer of a gamma phosphate from adenosinetriphosphate to a peptide or protein substrate). In certain embodiments,provided are methods for identifying a candidate molecule that interactswith a protein kinase, which comprise: contacting a compositioncontaining a protein kinase and a compound described herein with acandidate molecule under conditions in which the compound and theprotein kinase interact, and determining whether the amount of thecompound that interacts with the protein kinase is modulated relative toa control interaction between the compound and the protein kinasewithout the candidate molecule, whereby a candidate molecule thatmodulates the amount of the compound interacting with the protein kinaserelative to the control interaction is identified as a candidatemolecule that interacts with the protein kinase. Systems in suchembodiments can be a cell-free system or a system comprising cells(e.g., in vitro). The protein kinase, the compound or the molecule insome embodiments is in association with a solid phase. In certainembodiments, the interaction between the compound and the protein kinaseis detected via a detectable label, where in some embodiments theprotein kinase comprises a detectable label and in certain embodimentsthe compound comprises a detectable label. The interaction between thecompound and the protein kinase sometimes is detected without adetectable label.

Provided also are compositions of matter comprising a protein kinase anda compound described herein. In certain embodiments, the compound in thecomposition is not compound A2, compound A1 or compound A3. In someembodiments, the protein kinase in the composition is a serine-threonineprotein kinase or a tyrosine protein kinase. In certain embodiments, theprotein kinase is a protein kinase fragment having compound-bindingactivity. In some embodiments, the protein kinase in the composition is,or contains a subunit (e.g., catalytic subunit, SH2 domain, SH3 domain)of, CK2, Pim subfamily protein kinase (e.g., PIM1, PIM2, PIM3) or Fltsubfamily protein kinase (e.g, FLT1, FLT3, FLT4). In certain embodimentsthe composition is cell free and sometimes the protein kinase is arecombinant protein.

The protein kinase can be from any source, such as cells from a mammal,ape or human, for example. Examples of serine-threonine protein kinasesthat can be inhibited, or may potentially be inhibited, by compoundsdisclosed herein include without limitation human versions of CK2,CK2a2, Pim subfamily kinases (e.g., PIM1, PIM2, PIM3), CDK1/cyclinB,c-RAF, Mer, MELK, HIPK3, HIPK2 and ZIPK. A serine-threonine proteinkinase sometimes is a member of a sub-family containing one or more ofthe following amino acids at positions corresponding to those listed inhuman CK2: leucine at position 45, methionine at position 163 andisoleucine at position 174. Examples of such protein kinases includewithout limitation human versions of CK2, STK10, HIPK2, HIPK3, DAPK3,DYK2 and PIM-1. Examples of tyrosine protein kinases that can beinhibited, or may potentially be inhibited, by compounds disclosedherein include without limitation human versions of Flt subfamilymembers (e.g., FLT1, FLT2, FLT3, FLT3 (D835Y), FLT4). An example of adual specificity protein kinase that can be inhibited, or maypotentially be inhibited, by compounds disclosed herein includes withoutlimitation DYRK2. Nucleotide and amino acid sequences for proteinkinases and reagents are publicly available (e.g., World Wide Web URLsncbi.nlm.nih.gov/sites/entrez/ and Tnvitrogen.com). For example, variousnucleotide sequences can be accessed using the following accessionnumbers: NM_(—)002648.2 and NP_(—)002639.1 for PIM1; NM_(—)006875.2 andNP_(—)006866.2 for PIM2; XM_(—)938171.2 and XP_(—)943264.2 for PIM3;NM_(—)004119.2 and NP_(—)004110.2 for FLT3; NM_(—)002020.3 andNP_(—)002011.2 for FLT4; and NM_(—)002019.3 and NP_(—)002010.2 for FLT1.

The invention also in part provides methods for treating a conditionrelated to aberrant cell proliferation. For example, provided aremethods of treating a cell proliferative condition in a subject, whichcomprises administering a compound described herein to a subject in needthereof in an amount effective to treat the cell proliferativecondition. The subject may be a research animal (e.g., rodent, dog, cat,monkey), optionally containing a tumor such as a xenograft tumor (e.g.,human tumor), for example, or may be a human. A cell proliferativecondition sometimes is a tumor or non-tumor cancer, including but notlimited to, cancers of the colorectum, breast, lung, liver, pancreas,lymph node, colon, prostate, brain, head and neck, skin, liver, kidney,blood and heart (e.g., leukemia, lymphoma, carcinoma). In someembodiments, the cell proliferative condition is a non-tumor cancer. Insome such embodiments, the non-tumor cancer is a hematopoietic cancer.In specific embodiments, it is acute myelogenous leukemia. In some suchembodiments, the leukemia is refractory AML or wherein the AML isassociated with a mutated Flt3.

Also provided are methods for treating a condition related toinflammation or pain. For example, provided are methods of treating painin a subject, which comprise administering a compound described hereinto a subject in need thereof in an amount effective to treat the pain.Provided also are methods of treating inflammation in a subject, whichcomprises administering a compound described herein to a subject in needthereof in an amount effective to treat the inflammation. The subjectmay be a research animal (e.g., rodent, dog, cat, monkey), for example,or may be a human.

Conditions associated with inflanunation and pain include, withoutlimitation, acid reflux, heartburn, acne, allergies and sensitivities,Alzheimer's disease, asthma, atherosclerosis, bronchitis, carditis,celiac disease, chronic pain, Crohn's disease, cirrhosis, colitis,dementia, dermatitis, diabetes, dry eyes, edema, emphysema, eczema,fibromyalgia, gastroenteritis, gingivitis, heart disease, hepatitis,high blood pressure, insulin resistance, interstitial cystitis, jointpain/arthritis/rheumatoid arthritis, metabolic syndrome (syndrome X),myositis, nephritis, obesity, osteopenia, glomerulonephritis (GN),juvenile cystic kidney disease, and type I nephronophthisis (NPHP),osteoporosis, Parkinson's disease, Guam-Parkinson dementia, supranuclearpalsy, Kuf's disease, and Pick's disease, as well as memory impairment,brain ischemia, and schizophrenia, periodontal disease, polyarteritis,polychondritis, psoriasis, scleroderma, sinusitis, Sjögren's syndrome,spastic colon, systemic candidiasis, tendonitis, urinary trackinfections, vaginitis, inflammatory cancer (e.g., inflammatory breastcancer) and the like.

Methods for determining effects of compounds herein on pain orinflammation are known. For example, formalin-stimulated pain behaviorsin research animals can be monitored after administration of a compounddescribed herein to assess treatment of pain (e.g., Li et al., Pain115(1-2): 182-90 (2005)). Also, modulation of pro-inflammatory molecules(e.g., IL-8, GRO-alpha, MCP-1, TNFalpha and iNOS) can be monitored afteradministration of a compound described herein to assess treatment ofinflammation (e.g., Parhar et al., Int J Colorectal Dis. 22(6): 601-9(2006)), for example. Thus, also provided are methods for determiningwhether a compound herein reduces inflammation or pain, which comprisecontacting a system with a compound described herein in an amounteffective for modulating (e.g., inhibiting) the activity of a painsignal or inflammation signal.

Provided also are methods for identifying a compound that reducesinflammation or pain,

which comprise: contacting a system with a compound of one of theFormulae described herein, including a compound of Formula IA, IB, IC,L, L-A or L-B, and detecting a pain signal or inflammation signal,whereby a compound that modulates the pain signal relative to a controlmolecule is identified as a compound that reduces inflammation of pain.Non-limiting examples of pain signals are formalin-stimulated painbehaviors and examples of inflammation signals include withoutlimitation a level of a pro-inflammatory molecule.

The invention thus in part pertains to methods for modulatingangiogenesis in a subject, and methods for treating a conditionassociated with aberrant angiogenesis in a subject. proliferativediabetic retinopathy.

CK2 has also been shown to play a role in the pathogenesis ofatherosclerosis, and may prevent atherogenesis by maintaining laminarshear stress flow. CK2 plays a role in vascularization, and has beenshown to mediate the hypoxia-induced activation of histone deacetylases(HDACs). CK2 is also involved in diseases relating to skeletal muscleand bone tissue, including, e.g., cardiomyocyte hypertrophy, heartfailure, impaired insulin signaling and insulin resistance,hypophosphatemia and inadequate bone matrix mineralization.

Thus in one aspect, the invention provides methods to treat theseconditions, comprising administering to a subject in need of suchtreatment an effect amount of a CK2 inhibitor, such as a compound ofFormula A.

Thus, provided are methods for determining whether a compound hereinmodulates angiogenesis, which comprise contacting a system with acompound described herein in an amount effective for modulating (e.g.,inhibiting) angiogenesis or a signal associated with angiogenesis.Signals associated with angiogenesis are levels of a pro-angiogenesisgrowth factor such as VEGF. Methods for assessing modulation ofangiogenesis also are known, such as analyzing human endothelial tubeformation (BD BioCoat™ Angiogenesis System from BD Biosciences).Provided also are methods for identifying a compound that modulatesangiogenesis, which comprise contacting a system with a compound of oneof the Formulae described herein, including a compound of Formulae IA,IB, IC, L, L-A or L-B; and detecting angiogenesis in the system or anangiogenesis signal, whereby a compound that modulates the angiogenesisor angiogenesis signal relative to a control molecule is identified as acompound that modulates angiogenesis. Also provided are methods fortreating an angiogenesis condition, which comprise administering acompound described herein to a subject in need thereof in an amounteffective to treat the angiogenesis condition. Angiogenesis conditionsinclude without limitation solid tumor cancers, varicose disease and thelike.

The invention also in part pertains to methods for modulating an immuneresponse in a subject, and methods for treating a condition associatedwith an aberrant immune response in a subject. Thus, provided aremethods for determining whether a compound herein modulates an immuneresponse, which comprise contacting a system with a compound describedherein in an amount effective for modulating (e.g., inhibiting) animmune response or a signal associated with an immune response. Signalsassociated with immunomodulatory activity include, e.g., stimulation ofT-cell proliferation, suppression or induction of cytokines, including,e.g., interleukins, interferon-γ and TNF. Methods of assessingimmunomodulatory activity are known in the art. Provided also aremethods for identifying a compound that modulates an immune response,which comprise contacting a system with a compound of one of theFormulae described herein, including a compound of Formulae IA, IB, IC,L, L-A or L-B, or a pharmaceutically acceptable salt thereof; anddetecting immunomodulatory activity in a system, or a signal associatedwith immunomodulatory activity, whereby a compound that modulates theimmune response relative to a control molecule is identified as animmune response modulatory compound.

Also provided are methods for treating a condition associated with anaberrant immune response in a subject, which comprise administering acompound described herein to a subject in need thereof in an amounteffective to treat the condition. Conditions characterized by anaberrant immune response include without limitation, organ transplantrejection, asthma, autoimmune disorders, including rheumatoid arthritis,multiple sclerosis, my asthenia gravis, systemic lupus erythematosus,scleroderma, polymyositis, mixed connective tissue disease (MCTD),Crohn's disease, and ulcerative colitis. In certain embodiments, animmune response may be modulated by administering a compound herein incombination with a molecule that modulates (e.g., inhibits) thebiological activity of an mTOR pathway member or member of a relatedpathway (e.g., mTOR, PI3 kinase, AKT). In certain embodiments themolecule that modulates the biological activity of an mTOR pathwaymember or member of a related pathway is rapamycin. In certainembodiments, provided herein is a composition comprising a compounddescribed herein in combination with a molecule that modulates thebiological activity of an mTOR pathway member or member of a relatedpathway, such as rapamycin, for example.

In preferred embodiments of the present invention, the compound is acompound of Formula IA, IB, IC, L, L-A or L-B in one of the Tablesprovided herein, or a pharmaceutically acceptable salt of one of thesecompounds.

Any suitable formulation of a compound described above can be preparedfor administration. Any suitable route of administration may be used,including, but not limited to, oral, parenteral, intravenous,intramuscular, transdermal, topical and subcutaneous routes. Dependingon the subject to be treated, the mode of administration, and the typeof treatment desired—e.g., prevention, prophylaxis, therapy; thecompounds are formulated in ways consonant with these parameters.Preparation of suitable formulations for each route of administrationare known in the art. A summary of such formulation methods andtechniques is found in Remington's Pharmaceutical Sciences, latestedition, Mack Publishing Co., Easton, Pa., which is incorporated hereinby reference. The formulation of each substance or of the combination oftwo substances will generally include a diluent as well as, in somecases, adjuvants, buffers, preservatives and the like. The substances tobe administered can be administered also in liposomal compositions or asmicroemulsions.

For injection, formulations can be prepared in conventional forms asliquid solutions or suspensions or as solid forms suitable for solutionor suspension in liquid prior to injection or as emulsions. Suitableexcipients include, for example, water, saline, dextrose, glycerol andthe like. Such compositions may also contain amounts of nontoxicauxiliary substances such as wetting or emulsifying agents, pH bufferingagents and the like, such as, for example, sodium acetate, sorbitanmonolaurate, and so forth.

Various sustained release systems for drugs have also been devised, andcan be applied to compounds of the invention. See, for example, U.S.Pat. No. 5,624,677, the methods of which are incorporated herein byreference.

Systemic administration may also include relatively noninvasive methodssuch as the use of suppositories, transdermal patches, transmucosaldelivery and intranasal administration. Oral administration is alsosuitable for compounds of the invention. Suitable forms include syrups,capsules, tablets, as is understood in the art.

For administration to animal or human subjects, the appropriate dosageof the a compound described above often is 0.01-15 mg/kg, and sometimes0.1-10 mg/kg. Dosage levels are dependent on the nature of thecondition, drug efficacy, the condition of the patient, the judgment ofthe practitioner, and the frequency and mode of administration; however,optimization of such parameters is within the ordinary level of skill inthe art.

Therapeutic Combinations

Compounds of the invention may be used alone or in combination withanother therapeutic agent. The invention provides methods to treatconditions such as cancer, inflammation and immune disorders byadministering to a subject in need of such treatment a therapeuticallyeffective amount of a therapeutic agent useful for treating saiddisorder and administering to the same subject a a therapeuticallyeffective amount of a modulator of the present invention. A CK2, Pim orFlt modulator is an agent that inhibits or enhances a biologicalactivity of a CK2 protein, a Pim protein or a Flt protein, and isgenerically referred to hereafter as a “modulator.” The therapeuticagent and the modulator may be administered together, either as separatepharmaceutical compositions or admixed in a single pharmaceuticalcomposition. The therapeutic agent and the modulator may also beadministered separately, including at different times and with differentfrequencies. The modulator may be administered by any known route, suchas orally, intravenously, intramuscularly, nasally, and the like; andthe therapeutic agent may also be administered by any conventionalroute. In many embodiments, at least one and optionally both of themodulator and the therapeutic agent may be administered orally.

When used in combination, in some embodiments the compounds of theinvention may be administered as a single pharmaceutical dosageformulation that contains both a compound of the invention and anothertherapeutic agent. In other embodiments, separate dosage formulationsare administered; the compound of the invention and the othertherapeutic agent may be, administered at essentially the same time, forexample, concurrently, or at separately staggered times, for example,sequentially. In certain examples, the individual components of thecombination may be administered separately, at different times duringthe course of therapy, or concurrently, in divided or single combinationforms. The present invention provides, for example, simultaneous,staggered, or alternating treatment. Thus, the compound of the inventionmay be administered at the same time as another therapeutic agent, inthe same pharmaceutical composition; the compound of the invention maybe administered in separate pharmaceutical compositions; the compound ofthe invention may be administered before the other therapeutic agent, orthe other therapeutic agent may be administered before the compound ofthe invention, for example, with a time difference of seconds, minutes,hours, days, or weeks. In examples of a staggered treatment, a course oftherapy with the compound of the invention may be administered, followedby a course of therapy with the other therapeutic agent, or the reverseorder of treatment may be used, more than one series of treatments witheach component may be used. In certain examples of the presentinvention, one component, for example, the compound of the invention orthe other therapeutic agent, is administered to a mammal while the othercomponent, or its derivative products, remains in the bloodstream of themammal. In other examples, the second component is administered afterall, or most of the first component, or its derivatives, have left thebloodstream of the mammal.

Compounds of the invention are useful when used in combination withalkylating agents, angiogenesis inhibitors, antibodies, antimetabolites,antimitotics, antiproliferatives, aurora kinase inhibitors, Bcr-Ablkinase inhibitors, biologic response modifiers, cyclin-dependent kinaseinhibitors, cell cycle inhibitors, cyclooxygenase-2 inhibitors, leukemiaviral oncogene homolog (ErbB2) receptor inhibitors, growth factorinhibitors, heat shock protein (HSP)-90 inhibitors, histone deacetylase(HDAC) inhibitors, hormonal therapies, immunologicals, intercalatingantibiotics, kinase inhibitors, mammalian target of rapomycininhibitors, mitogen-activated extracellular signal-regulated kinaseinhibitors, non-steroidal anti-inflammatory drugs (NSAID's), platinumchemotherapeutics, polo-like kinase inhibitors, proteasome inhibitors,purine analogs, pyrimidine analogs, receptor tyrosine kinase inhibitors,retinoids/deltoids, plant alkaloids, topoisomerase inhibitors and thelike.

Compounds of the invention are useful when used in combination withalkylating agents, angiogenesis inhibitors, antibodies, antimetabolites,antimitotics, antiproliferatives, aurora kinase inhibitors, Bcr-Ablkinase inhibitors, biologic response modifiers, cyclin-dependent kinaseinhibitors, cell cycle inhibitors, cyclooxygenase-2 inhibitors, leukemiaviral oncogene homolog (ErbB2) receptor inhibitors, growth factorinhibitors, heat shock protein (HSP)-90 inhibitors, histone deacetylase(HDAC) inhibitors, hormonal therapies, immunologicals, intercalatingantibiotics, kinase inhibitors, mammalian target of rapomycininhibitors, mitogen-activated extracellular signal-regulated kinaseinhibitors, non-steroidal anti-inflammatory drugs (NSAID's), platinumchemotherapeutics, polo-like kinase inhibitors, proteasome inhibitors,purine analogs, pyrimidine analogs, receptor tyrosine kinase inhibitors,retinoids/deltoids, plant alkaloids, topoisomerase inhibitors and thelike.

Alkylating agents include altretamine, AMD-473, AP-5280, apaziquone,bendamustine, brostallicin, busulfan, carboquone, carmustine (BCND),chlorambucil, VNP 40101M, cyclophosphamide, decarbazine, estramustine,fotemustine, glufosfamide, ifosfamide, KW-2170, lomustine (CCNU),mafosfamide, melphalan, mitobronitol, mitolactol, nimustine, nitrogenmustard N-oxide, ranimustine, temozolomide, thiotepa, treosulfan,trofosfamide and the like.

Angiogenesis inhibitors include endothelial-specific receptor tyrosinekinase (Tie-2) inhibitors, epidermal growth factor receptor (EGFR)inhibitors, insulin growth factor-2 receptor (TGFR-2) inhibitors, matrixmetalloproteinase-2 (MMP-2) inhibitors, matrix metalloproteinase-9(MMP-9) inhibitors, platelet-derived growth factor receptor (PDGFR)inhibitors, thrombospondin analogs vascular endothelial growth factorreceptor tyrosine kinase (VEGFR) inhibitors and the like.

Aurora kinase inhibitors include AZD-1152, MLN-8054, VX-680 and thelike.

Bcr-Abl kinase inhibitors include BMS-354825, imatinib and the like.

CDK inhibitors include AZD-5438, BMI-1040, BMS-032, BMS-387, CVT-2584,flavopyridol, GPC-286199, MCS-5A, PD0332991, PHA-690509, seliciclib(CYC202, R-roscovitine), ZK-304709 and the like.

COX-2 inhibitors include ABT-963, etoricoxib, valdecoxib, BMS347070,celecoxib, COX-189 (lumiracoxib), CT-3, deracoxib, JTE-522,4-methyl-2-(3,4-dimethylphenyl)-1-(4-sulfamoylphenyl-1H-pyrrole), MK-663(etoricoxib), NS-398, parecoxib, RS-57067, SC-58125, SD-8381, SVT-2016,S-2474, T-614, rofecoxib and the like.

EGFR inhibitors include ABX-EGF, anti-EGFr immunoliposomes, EGFvaccine,EMD-7200, cetuximab, HR3, IgA antibodies, gefitinib, erlotinib, TP-38,EGFR fusion protein, (lapatinib and the like.

ErbB2 receptor inhibitors include CP-724-714, CI-1033 (canertinib),trastuzumab, lapatinib, pertuzumab, TAK-165, GW-572016 (ionafarnib),GW-282974, EKB-569, PI-166, dHER2 (HER2 vaccine), APC-8024 (HER-2vaccine), anti-HER/2neu bi specific antibody, B7.her2IgG3, AS HER2trifunctional bispecfic antibodies, mAB AR-209, mAB 2B-1 and the like.

Histone deacetylase inhibitors include depsipeptide, LAQ-824, MS-275,trapoxin, suberoylanilide hydroxamic acid (SAHA), TSA, valproic acid andthe like.

HSP-90 inhibitors include 17-AAG-nab, 17-AAG, CNF-101, CNF-1010,CNF-2024, 17-DMAG, geldanamycin, IPI-504, KOS-953, MYCOGRAB®,NCS-683664, PU24FCI, PU-3, radicicol, SNX-2112, STA-9090, VER49009 andthe like.

MEK inhibitors include ARRY-142886, ARRY-438162, PD-325901, PD-98059 andthe like.

mTOR inhibitors include AP-23573, CC1-779, everolimus, RAD-001,rapamycin, temsirolimus and the like.

Non-steroidal anti-inflammatory drugs include salsalate, diflunisal,ibuprofen, ketoprofen, nabumetone, piroxicam, ibuprofin cream, naproxen,diclofenac, indomethacin, sulindac, tolmetin, etodolac, ketorolac,oxaprozin and the like.

PDGFR inhibitors include C-451, CP-673, CP-868596 and the like.

Platinum chemotherapeutics include cisplatin, oxaliplatin, eptaplatin,lobaplatin, nedaplatin, carboplatin, satraplatin and the like.

Polo-like kinase inhibitors include B1-2536 and the like.

Thrombospondin analogs include ABT-510, ABT-567, ABT-898, TSP-1 and thelike.

VEGFR inhibitors include bevacizumab, ABT-869, AEE-788, RPI.4610,axitinib (AG-13736), AZD-2171, CP-547,632, 1M-862, pegaptanib,sorafenib, pazopanib, PTK-787/ZK-222584, sunitinib, VEGF trap,vatalanib, vandetanib and the like.

Antimetabolites include pemetrexed, 5-azacitidine, capecitabine,carmofur, cladribine, clofarabine, cytarabine, cytosine arabinoside,decitabine, deferoxamine, doxifluridine, eflornithine, EICAR,enocitabine, ethnylcytidine, fludarabine, hydroxyurea, 5-fluorouracil(5-FU) alone or in combination with leucovorin, gemcitabine,hydroxyurea, melphalan, mercaptopurine, 6-mercaptopurine riboside,methotrexate, mycophenolic acid, nelarabine, nolatrexed, ocfosate,pelitrexol, pentostatin, raltitrexed, Ribavirin, triapine, trimetrexate,S-1, tiazofurin, tegafur, TS-1, vidarabine, UFT and the like.

Antibiotics include intercalating antibiotics aclarubicin, actinomycinD, amrubicin, annamycin, adriamycin, bleomycin, daunorubicin,doxorubicin, liposomal doxorubicin, elsamitrucin, epirbucin, glarbuicin,idarubicin, mitomycin C, nemorubicin, neocarzinostatin, peplomycin,pirarubicin, rebeccamycin, stimalamer, streptozocin, valrubicin,zinostatin and the like.

Topoisomerase inhibitors include aclarubicin, 9-aminocamptothecin,amonafide, amsacrine, becatecarin, belotecan, BN-80915, irinotecan,camptothecin, dexrazoxine, diflomotecan, edotecarin, epirubicin,etoposide, ex atecan, 10-hydroxycamptothecin, gimatecan, lurtotecan,mitoxantrone, orathecin, pirarbucin, pixantrone, rubitecan, sobuzoxane,SN-38, tafluposide, topotecan and the like.

Antibodies include bevacizumab, CD40-specific antibodies, chTNT-1/B,denosumab, cetuximab, zanolimumab, IGF1R-specific antibodies,lintuzumab, edrecolomab, WX-G250, rituxirnab, ticilimumab, trastuzimaband the like.

Hormonal therapies include anastrozole, exemestane, arzoxifene,bicalutamide, cetrorelix, degarelix, deslorelin, trilostane,dexamethasone, flutamide, raloxifene, fadrozole, toremifene,fulvestrant, letrozole, formestane, glucocorticoids, doxercalciferol,lasofoxifene, leuprolide acetate, megesterol, mifepristone, nilutamide,tamoxifen citrate, abarelix, predisone, finasteride, rilostane,buserelin, triptorelin, luteinizing hormone releasing hormone (LHRH),vantas, trilostane, fosrelin (goserelin) and the like.

Deltoids and retinoids include seocalcitol (EB 1089, CB1093),lexacalcitrol (KH1060), fenretinide, aliretinoin, liposomal tretinoin,bexarotene, LGD-1550 and the like.

Plant alkaloids include, but are not limited to, vincristine,vinblastine, vindesine, vinorelbine and the like.

Proteasome inhibitors include bortezomib, MG132, NPI-0052, PR-171 andthe like.

Examples of immunological s include interferons and otherimmune-enhancing agents. Interferons include interferon alpha,interferon alpha-2a, interferon alpha-2b, interferon beta, interferongamma-1a, interferon gamma-1b, or interferon gamma-n1, combinationsthereof and the like.

Other agents include ALFAFERONE®, BAM-002, tasonermin, tositumomab,alemtuzumab, CTLA4 (cytotoxic lymphocyte antigen 4), decarbazine,denileukin, epratuzumab, lenograstim, lentinan, leukocyte alphainterferon, imiquimod, MDX-010, melanoma vaccine, mitumomab,molgramostim, gemtuzumab ozogamicin, filgrastim, OncoVAC-CL, oregovomab,pemtumomab (Y-muHMFG1), sipuleucel-T, sargaramostim, sizofilan,teceleukin, TheraCys® (BCG live), ubenimex, VIRULIZIN®, Z-100, WF-I0,aldesleukin, thymalfasin, daclizumab, Ibritumomab tiuxetan and the like.

Biological response modifiers are agents that modify defense mechanismsof living organisms or biological responses, such as survival, growth,or differentiation of tissue cells to direct them to have anti-tumoractivity and include include krestin, lentinan, sizofuran, picibanilPF-3512676 (CpG-8954), ubenimex and the like.

Pyrimidine analogs include cytarabine (ara C), cytosine arabinoside,doxifluridine, fludarabine, 5-FU (5-fluorouracil), floxuridine,gemcitabine, ratitrexed, triacetyluridine troxacitabine and the like.

Purine analogs include thioguanine and mercaptopurine.

Antimitotic agents include batabulin, epothilone D,N-(2-((4hydroxyphenyl)amino)pyridin-3-yl)-4-methoxybenzenesulfonamide,ixabepilone (BMS 247550), paclitaxel, docetaxel, PNUI00940 (109881),patupilone (epothilone B), XRP-9881, vinflunine, ZK-EPO and the like.

Compounds of the present invention are also intended to be used as aradiosensitizer that enhances the efficacy of radiotherapy. Examples ofradiotherapy include, but are not limited to, external beamradiotherapy, teletherapy, brachtherapy and sealed and unsealed sourceradiotherapy.

Additionally, compounds of the invention may be combined with otherchemotherapeutic agents such as ABI-007, ABT-100 (farnesyl transferaseinhibitor), lovastatin, poly I:poly CI2U, exisulind, pamidronic acid,arglabin, L-asparaginase, atamestane(1-methyl-3,17-dione-androsta-1,4-diene), tazarotne, AVE-8062, BEC2(mitumomab), cachectin or cachexin (tumor necrosis factor), canvaxin(vaccine), CeaVac™ (cancer vaccine), celmoleukin, histaminedihydrochloride, human papillomavirus vaccine, cyclophosphamide;doxorubicin; Vincristine; prednisone, Cyproterone Acetate, combrestatinA4P, DAB(389)EGF or TransMID-107R™ (diphtheria toxins), dacarbazine,dactinomycin, 5,6-dimethylxanthenone-4-acetic acid (DMXAA), eniluracil,squalamine lactate, T4N5 liposome lotion, discodermolide, DX-8951f(exatecan mesylate), enzastaurin, EP0906, quadrivalent humanpapillomavirus (Types 6, 11, 16, 18) recombinant vaccine, gastrimmune,genasense, GMK (ganglioside conjugate vaccine), GVAX® (prostate cancervaccine), halofuginone, hi sterelin, hydroxycarbamide, ibandronic acid,IGN-101, IL-13-PE38, IL-13-PE38QQR (cintredekin besudotox),IL-13-pseudomonas exotoxin, interferon-α, interferon-γ, mifamurtide,lonafamib, 5,10-methylenetetrahydrofolate, miltefosine(hexadecylphosphocholine), AE-941, trimetrexate glucuronate,pentostatin, ONCONASE® (a ribonuclease enzyme), ONCOPHAGE® (melanomavaccine treatment), OncoVAX (IL-2 Vaccine), rubitecan, OSIDEM®(antibody-based cell drug), OvaRex® MAb (murine monoclonal antibody),paclitaxel, aglycone saponins from ginseng comprising20(S)protopanaxadiol (aPPD) and 20(S)protopanaxatriol (aPPT)),panitumumab, PANVAC®-VF (investigational cancer vaccine), pegaspargase,PEG Interferon A, phenoxodiol, procarbazine, rebimastat, catumaxomab,lenalidomide, RSR13 (efaproxiral), lanreotide, acitretin, staurosporine(Streptomyces staurospores), talabostat (PTI00), bexarotene,DHA-paclitaxel, TLK286, temilifene, temozolomide, tesmilifene,thalidomide, STn-KLH, thymitaq(2-amino-3,4-dihydro-6-methyl-4-oxo-5-(4pyridylthio)-quinazolinedihydrochloride), TNFerade™ (adenovector: DNA carrier containing thegene for tumor necrosis factor-α), bosentan, tretinoin (Retin-A),tetrandrine, arsenic trioxide, VIRULIZIN®, ukrain (derivative ofalkaloids from the greater celandine plant), vitaxin (anti-alphavbeta3antibody), motexafin gadolinium, atrasentan, paclitaxel poliglumex,trabectedin, ZD-6126, dexrazoxane, zometa (zolendronic acid), zorubicinand the like.

In certain embodiments, a modulator compound of the invention may beused in combination with a therapeutic agent that can act by binding toregions of DNA that can form certain quadruplex structures. In suchembodiments, the therapeutic agents have anticancer activity on theirown, but their activity is enhanced when they are used in combinationwith a modulator. This synergistic effect allows the therapeutic agentto be administered in a lower dosage while achieving equivalent orhigher levels of at least one desired effect.

A modulator may be separately active for treating a cancer. Forcombination therapies described above, when used in combination with atherapeutic agent, the dosage of a modulator will frequently be two-foldto ten-fold lower than the dosage required when the modulator is usedalone to treat the same condition or subject. Determination of asuitable amount of the modulator for use in combination with atherapeutic agent is readily determined by methods known in the art.

The following examples illustrate and do not limit the invention.

Example 1 Processes for Synthesizing Compounds of Formulae I, II, IIIand IV Process 1

3-bromo-4-pyridine carboxylic acid (3.0 g, 14.9 mmol) in ethanol (100mL) was treated with concentrated sulfuric acid (5 mL).

The mixture was brought to reflux at which time everything went intosolution. After 12 hours at reflux, LCMS indicated that the reaction wascomplete. The reaction mixture was cooled to room temperature andconcentrated on a rotary evaporator to a third of its original volume.The mixture was then diluted with 250 mL of ethyl acetate and washedtwice with saturated aqueous sodium bicarbonate. Concentration on arotary evaporator yielded 3.25 g of the ethyl ester as a yellowish oilwhich was sufficiently pure enough for subsequent chemicaltransformations. LCMS (ESI) 216.2 (M+1)⁺.

Ethyl 3-bromo-4-pyridine carboxylate 1.15 g, 5.0 mmol),2-amino-4-methoxycarbonyl-phenylboronic acid (1.04 g, 4.5 mmol), sodiumacetate (1.64 g, 20 mmol), 1,1′-bis(diphenylphosphino)ferrocenepalladium (II) chloride (complexed with dichloromethane) (182 mg, 0.25mmol) and dimethylformamide (7.5 mL) were combined in a flask. The flaskwas evacuated and filled with nitrogen twice and heated to 125° C. withstirring for 12 hours or until LCMS indicated the absence of anystarting material. The mixture was cooled to room temperature and water(100 mL) was added to form a brown precipitate. The precipitate wasfiltered to yield 637 mg of methyl5-oxo-5,6-dihydrobenzo[c][2,6]naphthyridine-8-carboxylate. LCMS (EST)255.4 (M+1)⁺.

Methyl 5-oxo-5,6-dihydrobenzo[c][2,6]naphthyridine-8-carboxylate (200mg, 0.787 mmol) was combined with phosphorus oxychloride (1 mL) andheated to reflux. After 2 hours, LCMS indicated the absence of anystarting material. The volatiles were removed under reduced pressure.The residue was taken up in dichloromethane (50 mL) and washed twicewith saturated aqueous sodium bicarbonate. The organic phase was driedover sodium sulfate and concentrated on a rotary evaporator to givemethyl 5-chlorobenzo[c][2,6]naphthyridine-8-carboxylate (140 mg) as agrayish solid. LCMS (ESI) 273.3 (M+1)⁺.

Methyl 5-chlorobenzo[c][2,6]naphthyridine-8-carboxylate (20 mg, 0.074mmol) was combined with aniline (60 mg, 0.65 mmol) andN-methylpyrrolidinone (0.2 mL) in a microwave tube and the mixture washeated to 120° C. for 10 minutes at which time LCMS indicated that thereaction was complete as indicated by the absence of any startingmaterial. The mixture was then purified by HPLC to yield the ester (22mg) or it could be treated with 6N sodium hydroxide to yield the acid(19 mg). LCMS (ESI) 316.3 (M+1)⁺. ¹HNMR (400 MHz, CD₃OD) 10.17 (1H, s),9.67 (1H, br), 8.99 (1H, d, 5.9 Hz), 8.83 (1H, d, 8.6 Hz), 8.62 (1H, d,5.9 Hz), 8.24 (1H, d, 1.6 Hz), 8.04 (1H, s), 8.02 (1H, s), 7.93 (1H, dd,8.2, 1.6 Hz), 7.43 (1H, d, 7.4 Hz), 7.41 (1H, d, 7.4 Hz), 7.10 (1H, m).

Methyl 5-chlorobenzo[c][2,6]naphthyridine-8-carboxylate (232 mg, 0.853mmol) was combined with meta-chloroaniline (217 mg, 1.71 mmol) andN-methylpyrrolidinone (1 mL) in a flask and the mixture was heated to80° C. for 2 hours at which time LCMS indicated that the reaction wascomplete as indicated by the absence of any starting material. Themixture was dissolved in CH₂Cl₂, washed with saturated aqueous sodiumbicarbonate and dried over Na₂SO₄. The material was purified by flashchromatography (SiO₂, 1:1 to 9:1 gradient of EtOAc/Hexanes) to obtainthe ester. The material was dissolved in methanol and 6N aqueous NaOHand the mixture stirred at 50° C. for 30 minutes. The volatiles wereremoved in vacuo. The residue was triturated from aceticacid/THF/methanol using a mixture of hexanes and ethylacetate.Filtration and drying provided 147 mg of5-(3-chlorophenylamino)benzo[c][2,6]naphthyridine-8-carboxylic acid.LCMS (ESI) 350 (M+1)⁺. ¹HNMR (400 MHz, DMSO-d₆) δ 10.21 (s, 1H), 9.72(br s, 1H), 9.02 (d, J=5.6, 1H), 8.89 (d, J=8.8, 1H), 8.62 (d, J=5.6,1H), 8.31 (br s, 1H), 8.28 (d, J=1.6, 1H), 8.10 (br d, J=8, 1H), 7.99(dd, J=2, J=8.4, 1H), 7.46 (t, J=8.0, 1H), 7.16 (br d, J=7.2, 1H) ppm.

Sodium acetate (410 mg, 5 mmol) and 1,1′-bis(diphenylphosphino)ferrocenepalladium (II) chloride (complexed with dichloromethane) (36 mg, 0.05mmol) were added to a mixture of ethyl 3-bromo-4-pyridine carboxylate(230 mg, 1.0 mmol) and 2-amino-4-cyanophenylboronic acid hydrochloricacid salt (179 mg, 0.9 mmol). The mixture was connected to an exitbubbler and heated to 120° C. for 18 hours at which time LCMS analysisindicated that the reaction was done based on the disappearance ofstarting material. After cooling to room temperature, water was addedand the dark solids were filtered and washed with dichloromethane togive 5-oxo-5,6-dihydrobenzo[c][2,6]naphthyridine-8-carbonitrile (156 mg)as a gray solid which was sufficiently pure enough for subsequentchemical transformations. LCMS (ESI) 222.4 (M+1)⁺. ¹HNMR (400 MHz,DMSO-d₆) 12.2 (1H, s), 9.96 (1H, s), 8.90 (1H, d, 5.1 Hz), 8.77 (1H, d,8.2 Hz), 8.13 (1H, d, 5.1 Hz), 7.73 (1H, dd 8.2, 1.6 Hz), 7.70 (1H, d,1.6 Hz).

Phosphorus oxychloride (2 mL) was added to the5-oxo-5,6-dihydrobenzo[c][2,6]naphthyridine-8-carbonitrile (150 mg, 0.66mmol). The mixture was heated reflux for 3 hours at which time LCMSanalysis indicated the absence of any starting material. Volatiles wereremoved under vacuum and the crude product was dissolved indichloromethane, washed with brine and saturated aqueous sodiumbicarbonate and dried over sodium sulfate. After concentrating undervacuum, the crude product was triturated with ethyl acetate and hexanesto give 5-chlorobenzo[c][2,6]naphthyridine-8-carbonitrile (125 mg). LCMS(ESI) 240.3 (M+1)⁺.

A mixture of the 5-chlorobenzo[c][2,6]naphthyridine-8-carbonitrile (30mg, 0.13 mmol), aniline (60 mg, 0.65 mmol) and dimethylformamide (0.2mL) was heated to 120° C. in a microwave reactor for 10 minutes. LCMSindicated that absence of starting material. The mixture was dilutedwith water and left to stand for a few minutes as5-(phenylamino)benzo[c][2,6]naphthyridine-8-carbonitrile (25 mg)precipitated as an off-white solid. LCMS (ESI) 297.3 (M+1)⁺.

Sodium azide (65 mg, 1 mmol) and ammonium chloride (53 mg, 1 mmol) wereadded to a crude mixture of the5-(phenylamino)benzo[c][2,6]naphthyridine-8-carbonitrile (25 mg, 0.084mmol) in dimethylformamide (0.2 mL). The mixture was heated for 18 h at120° C. at which time LCMS analysis indicated the absence of anystarting material. The mixture was diluted with water and purified bypreparative HPLC to giveN-phenyl-8-(1H-tetrazol-5-yl)benzo[c][2,6]naphthyridin-5-amine (14 mg).LCMS (ESI) 340.3 (M+1)⁺. ¹HNMR (400 MHz, CD₃OD) 10.11 (1H, s), 8.96 (1H,d, 5.9 Hz), 8.85 (1H, d, 8.2 Hz), 8.53 (1H, d, 5.5 Hz), 8.47 (1H,$),8.16 (1H, d, 8.6 Hz), 7.88 (1H, s), 7.86 (1H, d, 0.8 Hz), 7.57-7.51 (3H,m), 7.36-7.31 (2H, m).

Representative compounds are set forth hereafter in Table 1.

TABLE 1 Molec- ular LCMS Compound Weight (ES) m/z

239.2 240 [M + 1]⁺

297.3 298 [M + 1]⁺

297.3 298 [M + 1]⁺

263.3 264 [M + 1]⁺

240.2 241 [M + 1]⁺

254.2 255 [M + 1]⁺

309.4 310 [M + 1]⁺

314.3 315 [M + 1]⁺

321.3 322 [M + 1]⁺

315.3 316 [M + 1]⁺

310.4 311 [M + 1]⁺

264.3 265 [M + 1]⁺

339.4 340 [M + 1]⁺

334.4 335 [M + 1]⁺

329.4 330 [M + 1]⁺

345.4 346 [M + 1]⁺

367.8 368 [M + 1]⁺ LCMS (ES) m/z Structure MW [M + 1]+

322.36 323

395.41 396

407.42 408

499.97 500

330.34 331

379.41 380

344.37 345

330.34 331

369.42 370

398.46 399

392.45 393

403.43 404

386.41 387

407.42 408

372.38 373

394.40 395

399.32 400

359.38 360

372.38 373

399.32 400

345.35 346

329.35 330

421.45 422

421.45 422

373.36 374

409.44 410

381.39 382

394.43 395

333.34 334

347.37 348

398.46 399

268.27 269

305.29 306

330.30 331

359.38 360

269.26 270

283.28 284

322.36 323

329.35 330

366.37 367

365.38 366

380.40 381

432.47 433

396.40 397

329.35 330

349.77 350

343.38 344

341.36 342

399.32 400

358.39 359

358.39 359

407.42 408

412.21 413

394.22 395

366.37 367

363.80 364

355.35 356

358.33 359

369.38 370

367.76 367

346.38 347

332.36 333

372.42 373

358.39 359

353.41 354

339.39 340

339.39 340

325.36 326

350.41 351

412.21 413

345.35 346

340.33 341

376.34 377

392.80 394

343.38 344

357.41 358

358.35 359

375.38 376

359.33 360

358.35 359

375.38 376

350.41 351

350.41 351

321.37 322

307.35 308

271.32 272

305.76 306

321.1 322

335.1 336

283.1 284

Process 2

5-bromopyrimidine-4-carboxylic acid (prepared according to the proceduredescribed in U.S. Pat. No. 4,110,450) (1.0 eq, 6.14 g, 30.2 mmol) wassuspended in CH₂Cl₂ (100 ml). Oxalylchloride (1.1 eq, 2.9 ml, 33.0 mmol)was added followed by 2 drops of DMF. The mixture was stirred at roomtemperature overnight and the volatiles were removed in vacuo. Theresidue was taken in MeOH (50 ml) and heated. After evaporation of MeOHin vacuo the compound was dissolved in CH₂Cl₂ and poured on a prepackedsilica gel column. The material was eluted using 20% Ethyl acetate inhexanes. Evaporation of the solvent providedmethyl-5-bromopyrimidine-4-carboxylate as a light orange crystallinesolid (2.54 g, 39% yield). LCMS (ES): 95% pure, m/z 217 [M]⁺; 219[M+2]⁺; ¹H NMR (CDCl₃, 400 MHz) δ 4.04 (s, 3H), 9.02 (s, 1H), 9.21 (s,1H) ppm.

Process 3

Sodium acetate (4.0 eq, 1.92 g, 23.41 mmol) and1,1′-bis(diphenylphosphino)ferrocene palladium (II) chloride (complexedwith dichloromethane) (0.05 eq, 214 mg, 0.29 mmol) were added to amixture of methyl-5-bromopyrimidine-4-carboxylate (1.0 eq, 1.27 g, 5.85mmol), and 2-amino-4-(methoxycarbonyl)phenylboronic acid hydrochloride(1.0 eq, 1.35 g, 5.85 mmol) in ahydrous DMF (10 ml). The Mixture wasstirred under nitrogen atmosphere at 120° C. for 18 hours. Water andbrine were added and the resulting solid impurities filtered off. Thematerial was extracted with CH₂Cl₂ (4×) and the combined extracts driedover Na₂SO₄. After evaporation of CH₂Cl₂, the remaining DMF wasevaporated by heating the residue in vacuo. The resulting solid wastriturated in CH₂Cl₂, filtered and dried to provide methyl5-oxo-5,6-dihydropyrimido[4,5-c]quinoline-8-carboxylate as a beige solid(127 mg, 8.5% yield). LCMS (ES): >80% pure, m/z 256 [M+1]⁺; ¹H NMR(DMSO-d₆, 400 MHz) δ 3.79 (s, 3H), 7.81 (d, J=8.0, 1H), 8.68 (d, J=8.8,1H), 9.49 (s, 1H), 10.19 (s, 1H), 12.37 (s, 1H) ppm.

Process 4

In a vial, methyl5-oxo-5,6-dihydropyrimido[4,5-c]quinoline-8-carboxylate (1.0 eq, 151 mg,0.59 mmol) was mixed in toluene (1 ml) with DIEA (1.5 eq, 155 ul, 0.89mmol) and POCl₃ (5 eq, 270 ul, 3.0 mmol). The mixture was stirred at120° C. for 1 hour and cooled down to room temperature. After adding iceand water the compound was extracted with CH₂Cl₂ (4×). The solution wasfiltered over Na₂SO₄ and filtered through a pad of celite. Afterevaporation of the volatiles, the material was triturated in a mixtureof ethyl acetate and hexanes, filtered and dried to afford methyl5-chloropyrimido[4,5-c]quinoline-8-carboxylate as a light brown fluffysolid (115 mg, 71% yield). LCMS (ES): 95% pure, m/z 274 [M+1]⁺. ¹H NMR(DMSO-d₆, 400 MHz) δ 3.96 (s, 3H), 8.37 (dd, J=1.6, J=8.4, 1H), 8.60 (d,J=1.6, 1H), 9.15 (d, J=8.8, 1H), 9.74 (s, 1H), 10.61 (s, 1H) ppm

Process 5

methyl 5-chloropyrimido[4,5-c]quinoline-8-carboxylate (10 mg) was mixedwith 3,5-difluoroaniline (100 mg) in NMP (0.1 ml). The mixture washeated under microwaves at 120° C. for 10 minutes. Water was added andthe material extracted with Cl₂Cl₂. The solvent was removed. Triturationin a mixture of ethylacetate and hexanes and filtration provided methyl5-(3,5-difluorophenylamino)pyrimido[4,5-c]quinoline-8-carboxylate. Thismaterial was suspended in a 1:1 mixture of THF and MeOH (2 ml) and a 5Naqueous solution of Lithium Hydroxide was added. The mixture wasvigorously stirred at room temperature for 5 hours. Water and 6Nhydrochloric acid were added to induce precipitation of the expectedmaterial. The solid was filtered, washed with water, dried and suspendedin MeOH. Filtration and drying gave5-(3,5-difluorophenylamino)pyrimido[4,5-c]quinoline-8-carboxylic acid asa yellow solid (4 mg, 31% yield). LCMS (ES): 95% pure, m/z 353 [M+1]⁺.¹H NMR (DMSO-d₆, 400 MHz) δ 6.90 (br t, J=9.6, 1H), 8.02 (dd, J=1.6,J=8.0, 1H), 8.18 (br d, J=10.8, 2H), 8.34 (d, J=1.6, 1H), 8.86 (d,J=8.4, 1H), 9.65 (s, 1H), 10.40 (s, 1H), 10.44 (s, 1H) ppm.

Process 6

5-(3-ethynylphenylamino)pyrimido[4,5-c]quinoline-8-carboxylic acid wasprepared using the same method, starting from methyl5-chloropyrimido[4,5-c]quinoline-8-carboxylate and 3-ethynylaniline.LCMS (ES): 95% pure, m/z 341 [M+1]⁺. ¹H NMR (DMSO-d₆, 400 MHz) δ 4.20(s, 1H), 7.19 (d, J=7.6, 1H), 7.42 (t, J=8.0, 1H), 7.99 (dd, J=1.6,J=8.4, 1H), 8.30 (d, J=1.6, 1H), 8.34 (dd, J=1.6, J=8.0, 1H), 8.49 (brs, 1H), 8.85 (d, J=8.8, 1H), 9.65 (s, 1H), 10.11 (s, 1H), 10.43 (s, 1H)ppm.

Representative analogs (Table 2) were prepared by the same method usingmethyl 5-chloropyrimido[4,5-c]quinoline-8-carboxylate and appropriateamines.

TABLE 2 Structure MW LCMS (ES) m/z

382.78 383 [M + 1]⁺

368.75 369 [M + 1]⁺

334.30 335 [M + 1]⁺

350.76 351 [M + 1]⁺

384.3114 385 [M + 1]⁺

339.3501 340 [M + 1]+

Process 7

methyl-5-bromo-2-(methylthio)pyrimidine-4-carboxylate was preparedaccording to the procedure used in process 2 for the preparation ofmethyl-5-bromopyrimidine-4-carboxylate. LCMS (ES): >90% pure, m/z 263[M]⁺, 265 [M+2]⁺; ¹H NMR (CDCl₃, 400 MHz) δ 2.59 (s, 3H), 4.00 (s, 3H),8.71 (s, 1H) ppm.

Process 8

Methyl-5-bromo-2-(methylthio)pyrimidine-4-carboxylate (1.0 eq, 661 mg,2.52 mmol) was dissolved in CH₂Cl₂ (10 ml). meta-chloro perbenzoic acid(m-cpba, 77% pure grade, 2.5 eq, 1.42 g, 6.34 mmol) was added and themixture was stirred at room temperature for 1 hour. To the resultingsuspension was added anhydrous THF (10 ml), methylamine hydrochloride(10 eq, 1.7 g, 25.18 mmol) and DIEA (10 eq, 4.3 ml, 24.69 mmol) and themixture stirred at room temperature overnight. The solvents were removedin vacuo prior to adding CH₂Cl₂ and a saturated aqueous sodiumbicarbonate solution. The two phases were decanted and two furtherCH₂Cl₂ extractions were carried out. The combined extracts were driedover Na₂SO₄ and the solvents evaporated. Purification by flashchromatography on silica gel (20-30% ethylacetate in hexanes) providedmethyl 5-bromo-2-(methylamino)pyrimidine-4-carboxylate as an off-whitesolid (461 mg, 75% yield). LCMS (ES): >95% pure, m/z 246 [M]⁺, 248[M+2]⁺.

Process 9

Sodium acetate (3.0 eq, 240 mg, 2.93 mmol) and1,1′-bis(diphenylphosphino)ferrocene palladium (II) chloride (complexedwith dichloromethane) (0.05 eq, 36 mg, 0.049 mmol) were added to amixture of methyl 5-bromo-2-(methylamino)pyrimidine-4-carboxylate (1.0eq, 240 mg, 0.975 mmol), and 2-amino-4-(methoxycarbonyl)phenylboronicacid hydrochloride (1.0 eq, 226 mg, 0.98 mmol) in ahydrous DMF (2 ml).The mixture was stirred under microwave heating at 120° C. for 10 min.Addition of water induced precipitation of the expected compound thatwas filtered and dried. methyl3-(methylamino)-5-oxo-5,6-dihydropyrimido[4,5-c]quinoline-8-carboxylate(57 mg, 21% yield). LCMS (ES): >80% pure, m/z 285 [M+1]⁺.

Process 10

3-(methylamino)-5-(phenylamino)pyrimido[4,5-c]quinoline-8-carboxylicacid was prepared usign methods described in process 3 and 4 startingfrom methyl3-(methylamino)-5-oxo-5,6-dihydropyrimido[4,5-c]quinoline-8-carboxylate.The final product was purified by flash chromatography and isolated as ayellow solid (0.35 mg). LCMS (ES): >95% pure, m/z 346 [M+1]⁺.

Process 11

In a microwave vessel, methyl5-bromo-2-(methylthio)pyrimidine-4-carboxylate (1.0 eq, 274 mg, 1.18mmol), 2-amino-4-(methoxycarbonyl)phenylboronic acid hydrochloride (1.2eq, 329 mg, 1.42 mmol), and sodium acetate (3.0 eq, 291 mg, 3.55 mmol)were mixed in anhydrous DMF (2 ml). The mixture was degassed by bubblingnitrogen gas in the solution for 10 min and the reaction heated undermicrowaves at 120° C. for 30 min. After cooling down the expectedmaterial crashed out of NMP. The solid was filtered, suspended in waterfiltered and dried. The material was triturated in AcOEt and filteredgive a yellow solid. The same procedure was repeated 9 times using thesame amounts of materials to provide methyl3-(methylthio)-5-oxo-5,6-dihydropyrimido[4,5-c]quinoline-8-carboxylate(283 mg, 10% yield). LCMS (ES): >95% pure, m/z 302 [M+1]⁺, ¹H NMR(DMSO-d₆, 400 MHz) δ 2.71 (s, 3H), 3.89 (s, 3H), 7.80 (dd, J=1.6, J=8.4,1H), 7.97 (d, J=1.6, 1H), 8.59 (d, J=8.8, 1H), 9.98 (s, 1H), 12.34 (s,1H) ppm.

Process 12

methyl3-(methylthio)-5-oxo-5,6-dihydropyrimido[4,5-c]quinoline-8-carboxylate(1.0 eq, 279 mg, 0.926 mmol) was suspended in toluene (2 ml). POCl₃ (2ml) and DIEA (0.5 ml) were added and the mixture stirred at 120° C. for5 hours. The volatiles were removed in vacuo and CH₂Cl₂ was added. Theorganic phase was washed with saturated aqueous sodium bicarbonate,washed with water and dried over Na₂SO₄. The solution was filteredthrough a pad of celite and the solvents removed in vacuo. The materialwas triturated in hexanes and AcOEt, filtered and dried to providemethyl 5-chloro-3-(methylthio)pyrimido[4,5-c]quinoline-8-carboxylate asa beige solid (184 mg, 63% yield). LCMS (ES): >95% pure, m/z 320 [M+1]⁺,322 [M+3]⁺.

Process 13

methyl 5-chloro-3-(methylthio)pyrimido[4,5-c]quinoline-8-carboxylate(1.0 eq, 182 mg, 0.57 mmol) was mixed with aniline (0.5 ml) in NMP (1ml). The mixture was heated under microwave for 10 minutes at 120° C.Water was added and the resulting solid was filtered and dried. Thecompound was triturated in EtOAc and hexanes and filtered to affordmethyl3-(methylthio)-5-(phenylamino)pyrimido[4,5-c]quinoline-8-carboxylate asa yellow solid. LCMS (ES): >95% pure, m/z 377 [M+1]⁺. This material wassuspended in CH₂Cl₂ (4 ml) and meta-chloroperbenzoic acid (77% pure, 2.5eq, 165 mg, 0.737 mmol) was added in small portions. After one hour, anadditional amount (100 mg) of mcpba was added and the mixture stirredfor 1.5 hours. After addition of more CH₂E1₂, the organic phase waswashed with water (4×), dried over Na₂SO₄ and the solution was filteredthrough a pad of silica gel, eluting with a MeOH/CH₂Cl₂ mixture. Afterevaporation of the solvents, methyl3-(methylsulfonyl)-5-(phenylamino)pyrimido[4,5-c]quinoline-8-carboxylatewas isolated as a yellow solid (166 mg, 72% yield). LCMS (ES): >95%pure, m/z 409 [M+1]⁺, (DMSO-d₆, 400 MHz) δ 3.77 (s, 3H), 3.93 (s, 3H),7.15 (t, J=7.2, 1H), 7.45 (t, J=7.6, 2H), 7.99 (dd, J=2.0, J=8.4, 1H),8.16 (d, J=7.6, 2H), 8.28 (d, J=2.0, 1H), 8.89 (d, J=8.8, 1H), 9.76 (s,1H), 10.61 (s, 1H) ppm.

Process 14

In a closed vial, methyl3-(methylsulfonyl)-5-(phenylamino)pyrimido[4,5-c]quinoline-8-carboxylate(1.0 eq, 62 mg, 0.152 mmol) was mixed with Methylamine hydrochloride(100 mg), DIEA (260 ul) in DMF (1 ml). The mixture was stirred at 60° C.for 40 min. Addition of water induced precipitation of methyl3-(methylamino)-5-(phenylamino)pyrimido[4,5-c]quinoline-8-carboxylatewhich was isolated by filtration. This material was suspended in a 1:1:1mixture of THF, MeOH and water (4 ml), and vigorously stirred at 60° C.in the presence of LiOH (200 mg) for 1.5 hours. Water aqueous HCl wereadded and to reach pH=1. The solid was filtered, dried and triturated inAcOEt/hexanes to provide3-(methylamino)-5-(phenylamino)pyrimido[4,5-c]quinoline-8-carboxylicacid as a yellow solid (40 mg, 74% yield). LCMS (ES): >95% pure, m/z 346[M+1]⁺.

The following analogs (table 3) were prepared using the same method.After purification by preparative HPLC and genevac evaporation thematerial were isolated as solids.

TABLE 3 Molecular Structure Weight LCMS (ES) m/z

371.39 372 [M + 1]⁺

373.41 374 [M + 1]+

389.41 390 [M + 1]+

375.38 376 [M + 1]+

389.41 390 [M + 1]+

414.46 415 [M + 1]+

430.50 431 [M + 1]+

444.49 445 [M + 1]+

458.51 459 [M + 1]+

395.41 396 [M + 1]⁺

397.43 398 [M + 1]⁺

413.43 414 [M + 1]⁺

438.48 439 [M + 1]⁺

482.53 483 [M + 1]⁺

369.38 370 [M + 1]⁺

405.84 406 [M + 1]⁺

428.36 429 [M + 1]⁺

379.80 380 [M + 1]⁺

393.83 394 [M + 1]⁺

365.77 366 [M + 1]⁺

407.85 408 [M + 1]⁺

439.39 440 [M + 1]⁺

393.83 397 [M + 1]⁺

397.79 398 [M + 1]⁺

383.76 384 [M + 1]⁺

423.83 424 [M + 1]⁺

441.84 442 [M + 1]⁺

427.46 428 [M + 1]⁺

441.48 442 [M + 1]⁺

455.51 456 [M + 1]⁺

439.47 440 [M + 1]⁺

409.44 410 [M + 1]⁺

366.76 367 [M + 1]⁺

399.40 400 [M + 1]⁺

450.88 451 [M + 1]⁺

450.94 451 [M + 1]⁺

436.85 437 [M + 1]⁺

437.84 438 [M + 1]⁺

436.91 437 [M + 1]⁺

324.33 325 [M + 1]⁺

335.36 336 [M + 1]⁺

385.42 386 [M + 1]⁺

371.39 372 [M + 1]⁺

407.37 408 [M + 1]⁺

389.38 390 [M + 1]⁺

401.42 402 [M + 1]⁺

386.41 387 [M + 1]⁺

385.42 386 [M + 1]⁺

365.39 366 [M + 1]⁺

454.88 455 [M + 1]⁺

523.00 524 [M + 1]⁺

474.87 475 [M + 1]⁺

471.87 472 [M + 1]⁺

463.85 464 [M + 1]⁺

474.87 475 [M + 1]⁺

474.87 475 [M + 1]⁺

407.42 408 [M + 1]⁺

340.40 341 [M + 1]⁺

366.42 367 [M + 1]⁺

295.30 296 [M + 1]⁺

337.38 338 [M + 1]⁺

309.32 310 [M + 1]⁺

323.35 324 [M + 1]⁺

399.33 400 [M + 1]⁺

386.41 387 [M + 1]⁺

339.35 340 [M + 1]⁺

386.41 387 [M + 1]⁺

399.45 400 [M + 1]⁺

337.38 338 [M + 1]⁺

439.39 440 [M + 1]⁺

386.41 387 [M + 1]⁺

405.84 406 [M + 1]⁺

407.37 408 [M + 1]⁺

353.38 354 [M + 1]⁺

408.45 409 [M + 1]⁺

367.40 368 [M + 1]⁺

399.45 400 [M + 1]⁺

395.45 396 [M + 1]⁺

379.41 380 [M + 1]⁺

381.43 382 [M + 1]⁺ LCMS (ES) Structure MW m/z [M + 1]+

415.44 416

349.39 350

381.43 382

354.36 355

378.43 379

300.34 301

407.37 408

407.37 408

389.38 390

401.42 402

423.83 424

435.86 436

401.42 402

421.45 422

385.42 386

415.44 416

415.44 416

429.47 430

428.44 429

407.37 408

407.37 408

319.32 320

295.30 296

376.43 377

390.46 391

422.46 423

376.43 377

422.46 423

385.42 386

385.42 386

359.38 360

373.41 374

387.43 388

417.46 418

345.35 346

Process 15

3-(cyclopropylamino)-5-(3-(trifluoromethyl)phenylamino)pyrimido[4,5-c]quinoline-8-carboxylicacid (20 mg) was mixed with 2 equivalent of an appropriate primary aminein NMP (0.5 ml). HOBt (14 mg), triethylamine (13 uL) and EDCI (18 mg)were added and the mixture was stirred at 70° C. for 1 hour. Water andHCl were added and the material was isolated by filtration. Thisprotocol was used to prepare compounds shown in table 4

TABLE 4 Structure MW LCMS (ES) m/z

438.41 439 [M + 1]+

478.47 479 [M + 1]+

452.43 453 [M + 1]+ LCMS(ES)m/z, Structure MW [M + 1]+

538.52 539

339.35 340

348.79 349

362.81 363

376.84 377

388.85 389

427.50 428

334.38 335

348.40 349

374.44 375

425.49 426

392.45 393

410.47 411

447.53 448

377.44 378

362.43 363

424.88 425

439.90 440

461.94 462

439.90 440

406.86 407

438.91 439

431.92 432

445.94 446

459.93 460

431.90 432

454.91 455

418.88 419

439.90 440

518.01 519

391.85 392

353.38 354

379.41 380

367.40 368

393.44 394

409.44 410

429.47 430

454.48 455

424.50 425

436.51 437

398.46 399

424.50 425

412.49 413

398.46 399

412.49 413

424.50 425

366.78 368

366.78 368

382.34 383

378.81 380

Process 16

3-(cyclopropylamino)-5-(3-(trifluoromethyl)phenylamino)pyrimido[4,5-c]quinoline-8-carboxylicacid (100 mg, 0.23 mmol) was reacted with diphenylphosphoryl azide (50ul, 0.23 mmol) and triethylamine (34 ul, 0.23 mmol) in isopropanol (8ml). The mixture was stirred at 95° C. for 3 hours. The solvents wereremoved and the residue partitioned between water and ethylacetate. Theorganic layer was dried over Na₂SO₄ and the solvents removed in vacuo.Addition of CH₇Cl₂ induced formation of a solid that was filtered offand dried to afford isopropyl3-(cyclopropylamino)-5-(3-(trifluoromethyl)phenylamino)pyrimido[4,5-c]quinolin-8-ylcarbamate.LCMS (ES): 90% pure, m/z 497 [M+1]⁺.

To isopropyl 3-(cyclopropylamino)-5-(3-(trifluoromethyl)phenylamino)pyrimido[4,5-c]quinolin-8-ylcarbamate (60 mg) in EtOH (3 mL) was added1.5 mL of NaOH solution (6 N) and the mixture was heated at reflux for 3hrs. EtOH was removed and the residue obtained was partitioned betweendichloromethane and water. Organic layer was separated, washed withbrine and dried with sodium sulfate. Dichloromethane was removed and theyellow solid obtained was used in the next step without furtherpurification. 19.5 mg of the yellow solid was dissolved indichloromethane (3 mL) and acetyl chloride (7.4 μL) was added followedby triethyl amine (14.54 μL). The mixture was stirred at roomtemperature over night. Water and dichloromethane were added and organiclayer was isolated, washed with 1N NaOH, Brine, dried with sodiumsulfate and concentrate. The residue obtained was purified bypreparative TLC eluting with dichloromethane-methanol (9-1) to affordN-(3-(cyclopropylamino)-5-(3-(trifluoromethyl)phenylamino)pyrimido[4,5-c]quinolin-8-yl)acetamide.LCMS (ES) m/z 453 [M+1]+.

Example 2 Processes for Synthesizing Compounds of Formulae V, VI, VIIand VIII Process 1

2-bromo-3-thiophene carboxylic acid (1.0 eq, 12.56 g, 60.66 mmol) wassuspended in CH₂Cl₂ (200 ml). Oxalyl chloride (1.1 eq, 5.9 ml, 67.16mmol) and 5 drops of DMF were added, inducing formation of gas. Themixture was stirred overnight at room temperature and the volatiles wereremoved in vacuo. The resulting solid was suspended in dry methanol (150ml) and the mixture heated to ebullition. Evaporation of the solventsafforded methyl 2-bromo-3-thiophene carboxylate (13.16 g, 98% yield) asa crude brown oil. LCMS (ES): 99% pure, m/z not detected; ¹H NMR (CDCl₁,400 MHz) δ 3.88 (s, 3H), 7.23 (d, J=5.6, 1H), 7.56 (d, J=5.6, 1H) ppm.

Process 2

In a microwave vessel, methyl 2-bromo-3-thiophene carboxylate (1.0 eq,260 mg, 1.18 mmol), 2-amino-4-(methoxycarbonyl)phenylboronic acidhydrochloride (1.1 eq, 300 mg, 1.30 mmol), sodium acetate (3.0 eq, 292mg, 3.56 mmol) and PdCl₂(dppf) (0.05 eq, 31 mg, 0.059 mmol) were mixedtogether in anhydrous DMF (2 ml). The mixture was heated in a microwaveoven at 120° C. for 10 nm. Water was added and the solid filtered anddried. The material was suspended in CH₂Cl₂, filtered and dried toafford methyl 4-oxo-4,5-dihydrothieno[3,2-c]quinoline-7-carboxylate as ayellow solid (152 mg, 50% yield). LCMS (ES): 95% pure, m/z 260 [M+1]⁺;¹H NMR (CDCl₃, 400 MHz) δ 3.99 (s, 3H), 7.54 (d, J=5.2, 1H), 7.79 (d,J=4.8, 1H), 7.86 (d, J=8.4, 1H), 7.91 (dd, J=8.4, J=1.6, 1H), 8.03 (d,J=1.2, 1H) ppm.

Process 3

Methyl 4-oxo-4,5-dihydrothieno[3,2-c]quinoline-7-carboxylate (1.0 eq,618 mg, 2.38 mmol) was suspended in 10 ml of a mixture of MeOH, THF, andwater (1:1:1, v:v:v). LiOH (2.0 eq, 114 mg, 4.76 mmol) was added and themixture was stirred at room temperature for 2 hours. An additionalamount of LiOH (114 mg) was added and the mixture was stirred for anhour. LiOH (50 mg) was added and the mixture stirred for an additional 2hours. Water was added and the solution filtered through a pad ofcelite. The pad of celite was thoroughly washed with aqueous 1 N NaOH.The solution was acidified with 6 N aqueous HCl to induce precipitationof the expected material. Filtration and drying afforded4-oxo-4,5-dihydrothieno[3,2-c]quinoline-7-carboxylic acid as a yellowsolid (562 mg, 96% yield). LCMS (ES): 95% pure, m/z 246 [M+1]⁺; ¹H NMR(DMSO-d₆, 400 MHz) δ 7.61 (d, J=5.2, 1H), 7.73 (dd, J=1.6, J=8.0, 1H),7.88 (d, J=5.6, 1H), 7.92 (d, J=8.4, 1H), 8.02 (d, J=1.6, 1H), 11.92 (s,1H), 13.21 (br. s, 1H) ppm.

Process 4

4-oxo-4,5-dihydrothieno[3,2-c]quinoline-7-carboxylic acid (1.0 eq, 38mg, 0.155 mmol) was suspended in dioxane (1 ml). LiAlH₄ (7.0 eq, 40 mg,1.05 mmol) was added and the mixture stirred at 100° C. for 45 nm. Waterwas added, then MeOH and CH₂Cl₂. The solid salts were filtered off andwashed with MeOH and CH₂Cl₂. After evaporation of the volatiles invacuo, the material was dissolved in a mixture of NMP, MeOH and waterand was purified by preparative HPLC. Genevac evaporation afforded7-(hydroxymethyl)thieno[3,2-c]quinolin-4(5H)-one as an off-white solid(12 mg, 34%). LCMS (ES): 95% pure, m/z 232 [M+1]⁺; ¹H NMR (DMSO-d₆, 400MHz) δ 4.56 (s, 2H), 7.15 (d, J=7.6, 1H), 7.39 (br s, 1H), 7.55 (d,J=5.2, 1H), 7.73 (d, J=5.2, 1H), 7.76 (d, J=8.0, 1H), 11.73 (s, 1H) ppm.

Process 5

Methyl 4-oxo-4,5-dihydrothieno[3,2-c]quinoline-7-carboxylate (1.0 eq, 17mg, 0.066 mmol) was suspended in a mixture of chloroform (0.3 ml) andacetic acid (0.1 ml). NBS was added (9.5 eq, 112 mg, 0.63 mmol) and themixture stirred at 70° C. for 16 hours. Water and aqueous ammonia wasadded and the material was extracted with CH₂Cl₂ (2×). The combinedextracts were dried over Na₂SO₄ and the solvent removed in vacuo toprovide methyl2-bromo-4-oxo-4,5-dihydrothieno[3,2-c]quinoline-7-carboxylate (17 mg,76%). LCMS (ES): >85% pure, m/z 338 [M]⁺, 340 [M+2]⁺; ¹H NMR(CDCl₃/CD₃OD, δ: 1, 400 MHz) δ 3.99 (s, 3H), 7.30 (m, 1H), 7.69 (d,J=8.4, 1H), 7.45 (m, 1H), 7.88 (br d, J=8, 1H), 8.05 (br s, 1H) ppm.

Process 6

Methyl 2-bromo-4-oxo-4,5-dihydrothieno[3,2-c]quinoline-7-carboxylate(1.0 eq, 17 mg, 0.050 mmol) was suspended in a 1:1:1 mixture ofMeOH/THF/water (0.6 ml). LiOH (39 mg) was added and the mixture stirredat room temperature for one hour. Water and 6N HCl was added and theresulting precipitate was filtered. The material was purified bypreparative HPLC. Genevac evaporation provided2-bromo-4-oxo-4,5-dihydrothieno[3,2-c]quinoline-7-carboxylic acid as asolid (2.1 mg, 13% yield). LCMS (ES): >95% pure, m/z 324 [M]⁺,326[M+2]⁺; ¹H NMR (DMSO-d₆, 400 MHz) δ 7.75 (s, 1H), 7.75 (dd, J=1.6,J=8.0, 1H), 7.90 (d, J=8.4, 1H), 8.03 (d, J=1.6, 1H), 12.06 (s, 1H) ppm.

Process 7

In a closed vessel, Methyl4-oxo-4,5-dihydrothieno[3,2-c]quinoline-7-carboxylate (44 mg, 0.170mmol) was suspended in concentrated aqueous ammonia (1 ml). The mixturewas stirred at 100° C. overnight. Aqueous 1N NaOH was added and themixture stirred at room temperature for 2 hours. The solid was filteredand dried to provide4-oxo-4,5-dihydrothieno[3,2-c]quinoline-7-carboxamide as a brown solid(13 mg, 32% yield). LCMS (ES): 95% pure, m/z 245 [M+1]⁺¹.

Process 8

In a microwave vessel, methyl 2-bromo-3-thiophene carboxylate (1.0 eq,64 mg, 0.29 mmol), 2-amino phenyl boronic acid (1.2 eq, 48 mg, 0.35mmol), sodium acetate (3.0 eq, 71 mg, 0.86 mmol) and PdCl₂(dppf) (0.1eq, 15 mg, 0.028 mmol) were mixed together in anhydrous DMF (0.2 ml).The mixture was heated in a microwave oven at 120° C. for 5 nm. Thematerial was purified by preparative HPLC. Acetonitrile was evaporated,and the compound was extracted with CH₂Cl₂ (3×). The combined extractswere washed with water, dried over Na₂SO₄, and the solvents removed invacuo. Recrystallization in EtOH providedthieno[3,2-c]quinolin-4(5H)-one as a tan crystalline solid (7 mg, 12%yield). LCMS (ES): 95% pure, m/z 202 [M+1]⁺; ¹H NMR (CDCl₃/CD₃OD, δ: 1,400 MHz) δ 7.28 (m, 1H), 7.33 (m, 1H), 7.43-7.50 (m, 2H), 7.74 (d,J=4.4, 1H), 7.82 (d, J=7.6, 1H) ppm

Process 9

In a microwave vessel, methyl 2-bromo-3-thiophene carboxylate (1.0 eq,250 mg, 1.13 mmol), 2-amino-3-cyanophenyl boronic acid HCl (1.1 eq, 250mg, 1.24 mmol), sodium acetate (3.0 eq, 278 mg, 3.39 mmol) andPdCl₂(dppf) (0.007 eq, 4.3 mg, 0.0082 mmol) were mixed together inanhydrous DMF (2.5 ml). The mixture was heated in a microwave oven at120° C. for 10 nm. Water was added and the material extracted withCH₂Cl₂. The organic extracts were washed with brine, dried over Na₂SO₄and the solvents removed in vacuo. The resulting solid was sonicated inAcOEt, filtered and dried to afford4-oxo-4,5-dihydrothieno[3,2-c]quinoline-7-carbonitrile as a beige solid(121 mg, 48% yield). LCMS (ES): 95% pure, m/z 227 [M+1]⁺.

Process 10

4-oxo-4,5-dihydrothieno[3,2-c]quinoline-7-carbonitrile (1.0 eq, 20 mg,0.088 mmol) was dissolved in anhydrous DMF (0.15 ml). Sodium azide (4.0eq, 23 mg, 0.354 mmol) and ammonium chloride (4.0 eq, 19 mg, 0.354 mmol)were added and the mixture stirred at 120° C. overnight. The reactionmixture was cooled down and water was added. Addition of aqueous 6N HClinduced formation of a precipitate. After filtration and drying invacuo, 7-(1H-tetrazol-5-yl)thieno[3,2-c]quinolin-4(5H)-one was isolatedas a greenish solid (18 mg, 76% yield)). LCMS (ES): 95% pure, m/z 270[M+1]⁺, 242 [M+1−N₂]⁺; ¹H NMR (DMSO-d₆, 400 MHz) δ 7.64 (d, J=5.2, 1H),7.86 (dd, J=1.6, J=8.4, 1H), 7.89 (d, J=5.2, 1H), 8.09 (d, J=8.0, 1H),8.16 (d, J=1.6, 1H), 12.03 (s, 1H) ppm.

Process 11

Methyl 4-oxo-4,5-dihydrothieno[3,2-c]quinoline-7-carboxylate (1.0 eq, 18mg, 0.069 mmol) was dissolved in anhydrous DMF (0.4 ml). K₂CO₃ (7.0 eq,70 mg, 0.506 mmol) and 3-bromo-1-propanol (16 eq, 100 ul, 1.144 mmol)were added and the mixture stirred at 100° C. for 1.5 hour. After addingwater, the mixture was extracted with CH₂Cl₂. The combined extracts weredried over Na₂SO₄ and the solvents removed in vacuo. Compounds 8 and 9were separated by preparative TLC on silica gel (eluted twice with 30%AcOEt in hexanes, then once with 50% AcOEt in hexanes). The less polarcompound is methyl4-(3-hydroxypropoxy)thieno[3,2-c]quinoline-7-carboxylate (12 mg). LCMS(ES): 80% pure, m/z 318 [M+1]⁺. The more polar compound is methyl5-(3-hydroxypropyl)-4-oxo-4,5-dihydrothieno[3,2-c]quinoline-7-carboxylate(19 mg). LCMS (ES): 80% pure, m/z 318 [M+1]⁺. The two compounds wereused for the following step without any further purification.

Process 12

Methyl5-(3-hydroxypropyl)-4-oxo-4,5-dihydrothieno[3,2-c]quinoline-7-carboxylate(1.0 eq, 19 mg, 0.060 mmol) was dissolved in a 1:1:1 mixture of THF,MeOH and water (0.5 ml). LiOH (40 mg) was added and the resultingmixture stirred at room temperature for 1.5 hours. Water, MeOH and HClwere added and the solution purified by preparative HPLC. Genevacevaporation afforded5-(3-hydroxypropyl)-4-oxo-4,5-dihydrothieno[3,2-c]quinoline-7-carboxylicacid as a white solid (4 mg, 22% yield). LCMS (ES): 95% pure, m/z 304[M+1]⁺. ¹H NMR (CDCl₃/CD₃OD, δ: 1, 400 MHz) δ 2.08 (qi, J=6.0, 2H), 3.61(t, J=5.2, 2H), 4.62 (t, J=6.0, 2H), 7.53 (d, J=5.2, 1H), 7.77 (d,J=5.2, 1H), 7.93 (d, =8.0, 1H), 7.99 (dd, J=1.2, J=8.4, 1H), 8.26 (d,J=0.8, 1H) ppm.

Process 13

Methyl 4-(3-hydroxypropoxy)thieno[3,2-c]quinoline-7-carboxylate wasprepared according to the procedure used in process 12.4-(3-hydroxypropoxy)thieno[3,2-c]quinoline-7-carboxylic acid wasisolated as a solid (3 mg, 26% yield). LCMS (ES): 95% pure, m/z 304[M+1]⁺.

Process 14

Methyl5-(2-(dimethylamino)ethyl)-4-oxo-4,5-dihydrothieno[3,2-c]quinoline-7-carboxylatewas prepared according to the procedure used in process 11 starting frommethyl 4-oxo-4,5-dihydrothieno[3,2-c]quinoline-7-carboxylate and2-dimethylaminoethyl chloride. LCMS (ES): 95% pure, m/z 331 [M+1]⁺.

Process 15

5-(2-(dimethylamino)ethyl)-4-oxo-4,5-dihydrothieno[3,2-c]quinoline-7-carboxylicacid was prepared according to the procedure used in process 12.Preparative HPLC and genevac evaporation provided the material as a TFAsalt. LCMS (ES): 95% pure, m/z 317 [M+1]⁺, ¹H NMR (CDCl₃/CD₃OD, 9:1, 400MHz) δ 3.06 (s, 6H), 3.50 (t, J=7.6, 2H), 4.88 (t, J=7.6, 2H), 7.53 (d,J=5.2, 1H), 7.73 (d, J=5.6, 1H), 7.89 (d, J=8.4, 1H), 7.95 (br d, J=8.4,1H), 8.2 (br s, 1H) ppm.

Process 16

Methyl 4-oxo-4,5-dihydrothieno[3,2-c]quinoline-7-carboxylate (1.0 eq,1.50 g, 5.79 mmol) was suspended in dry toluene (15 ml). POCl₃ (1.2 eq,0.64 mmol, 6.99 mmol) and DIEA (0.8 eq, 0.81 mmol, 4.65 mmol) were addedand the mixture vigorously stirred at 120° C. for 3 hours under nitrogenatmosphere. The mixture was hydrolyzed by addition of ice and water. Thecompound was extracted with CH₂Cl₂ (4×). The combined extracts weredried over Na₂SO₄ and the black solution filtered through a pad ofcelite. After evaporation of the volatiles in vacuo, the resulting solidwas triturated in a mixture of AcOEt and hexanes. Filtration and dryingprovided methyl 4-chlorothieno[3,2-c]quinoline-7-carboxylate as a yellowfluffy solid (1.14 g, 71% yield). LCMS (ES): 95% pure, m/z 278 [M+1]⁺,¹H NMR (CDCl₃, 400 MHz) δ 4.01 (s, 3H), 7.72 (d, J=4.8, 1H), 7.74 (d,J=5.2, 1H), 8.14 (d, J=8.4, 1H), 8.25 (d, J=8.4, 1H), 8.85 (d, J=1.6,1H) ppm.

Process 17

4-chlorothieno[3,2-c]quinoline was prepared according to the procedureused in process 16, starting from thieno[3,2-c]quinolin-4(5H)-one.4-chlorothieno[3,2-c]quinoline was isolated as a solid (71 mg, 93%yield). LCMS (ES): 95% pure, m/z 220 [M+1]⁺, 223 [M+3]⁺.

Process 18

4-chlorothieno[3,2-c]quinoline-7-carbonitrile was prepared according tothe procedure used in process 16.4-chlorothieno[3,2-c]quinoline-7-carbonitrile was isolated as a yellowfluffy solid (833 mg, 77% yield). LCMS (ES): 95% pure, m/z 245 [M+1]⁺,247 [M+3]⁺.

Process 19

4-chlorothieno[3,2-c]quinoline-7-carbonitrile (1.0 eq, 23 mg, 0.094mmol), aniline (0.1 ml) and NMP (0.1 ml) were mixed in a vial. Themixture was heated in a microwave oven at 120° C. for 10 nm. Water wasadded and the resulting solid4-(phenylamino)thieno[3,2-c]quinoline-7-carbonitrile was filtered anddried. LCMS (ES): 95% pure, m/z 302 [M+1]⁺.

Process 20

4-(phenylamino)thieno[3,2-c]quinoline-7-carbonitrile (34 mg, 0.113 mmol)was dissolved in NMP (0.3 ml). 30% aqueous H₂O₂ (0.2 ml) was addedfollowed by addition of 6N NaOH (50 ul). The mixture was stirred at 50°C. for 2 hours. An extra amount of 30% aqueous H₂O₂ (0.3 ml) and 6N NaOH(100 ul) were added and a 70% conversion was achieved after 30 min.Water was added and the solid filtered and dried. The material wasfurther reacted under the same conditions in order to achieve a completetransformation. 4-(phenylamino)thieno[3,2-c]quinoline-7-carboxamide wasisolated as solid (30 mg, 83% yield). LCMS (ES): 95% pure, m/z 320[M+1]⁺.

Process 21

4-(phenylamino)thieno[3,2-c]quinoline-7-carboxamide (28 mg, 0.088 mmol)was suspended in N,N-dimethylformamide dimethylacetal and the mixturestirred at 80° C. under nitrogen atmosphere for 2 hours. The volatileswere removed in vacuo. Acetic acid (0.5 ml) and anhydrous hydrazine (0.1ml) and the mixture stirred at 115° C. for 1 hour. Water and brine wereadded and the solid filtered. The material was purified by preparativeHPLC. Genevac evaporation and trituration in AcOEt/hexanes affordedN-phenyl-7-(4H-1,2,4-triazol-3-yl)thieno[3,2-c]quinolin-4-amine as anoff-white solid (9 mg, 30% yield). LCMS (ES): 94% pure, m/z 344 [M+1]⁺.

Process 22

4-(phenylamino)thieno[3,2-c]quinoline-7-carbonitrile (1.0 eq, 27 mg,0.0897 mmol) and hydroxylamine hydrochloride (10 eq, 62 mg, 0.892 mmol)and K₂CO₃ (10 eq, 124 mg, 0.896 mmol) were mixed in EtOH (0.5 ml) andthe mixture heated under microwave at 100° C. for 10 min. The solid wereremoved by filtration and washed with EtOH. The solvents were removed invacuo. The crude material was suspended in chloroform (0.5 ml). Ethylchloroformate (20 ul) and triethylamine (20 ul) were added and themixture stirred at room temperature for 10 min. CH₂Cl₂ was added and theorganic phase was washed with brine. The organic phase was dried overNa₂SO₄ and the solvent removed. The crude material was suspended in NMP(1 ml) and heated under microwave at 160° C. for 10 min. The materialwas purified by preparative HPLC. Genevac evaporation afforded3-(4-(phenylamino)thieno[3,2-c]quinolin-7-yl)-1,2,4-oxadiazol-5(4H)-oneas an off-white solid (7 mg, 22% yield). LCMS (ES): 95% pure, m/z 361[M+1]⁺.

Process 23

4-chlorothieno[3,2-c]quinoline-7-carbonitrile (1.0 eq, 23 mg, 0.094mmol), aniline (0.1 ml) and NMP (0.1 ml) were mixed in a vial. Themixture was heated in a microwave oven at 120° C. for 10 nm. Water wasadded and the resulting solid4-(phenylamino)thieno[3,2-c]quinoline-7-carbonitrile was filtered anddried. LCMS (ES): 95% pure, m/z 302 [M+1]⁺. This material was mixed in avial with DMF (0.5 ml), NH₄Cl (50 mg) and NaN₃ (50 mg). The mixture wasstirred at 120° C. for 3 hours. After addition of water and filtration,N-phenyl-7-(1H-tetrazol-5-yl)thieno[3,2-c]quinolin-4-amine was isolatedas a beige solid (13 mg, 41% yield). LCMS (ES): 95% pure, m/z 345[M+1]⁺, 317 [M+1−N₂]⁺. ¹H NMR (DMSO-d₆, 400 MHz) δ 7.07 (t, J=7.2, 1H),7.40 (t, J=7.6, 2H), 8.00 (dd, J=1.6, J=8.4, 1H), 8.04 (d, J=5.2, 1H),8.10 (dd, J=1.2, J=8.8, 2H), 8.19 (d, J=8.0, 1H), 8.25 (d, J=5.6, 1H),8.43 (d, J=1.6, 1H), 9.34 (s, 1H) ppm.

Representative analogs (Table 5) were prepared by the same method using4-chlorothieno[3,2-c]quinoline-7-carbonitrile and appropriate amines.The reaction temperatures used for the microwave reactions ranged from120° C. to 180° C. After synthesis of the tetrazoles, the materials wereisolated by preparative HPLC/genevac evaporation. In some instances, thematerials precipitated after addition of water to the reaction mixtureand were isolated by filtration.

TABLE 5 Structure M.W. LCMS (ES) m/z

339.42 340 [M + 1]⁺

362.38 363 [M + 1]⁺

396.83 397 [M + 1]⁺

374.42 375 [M + 1]⁺

378.84 379 [M + 1]⁺

408.86 409 [M + 1]⁺

404.45 405 [M + 1]⁺

428.39 429 [M + 1]⁺

402.47 403 [M + 1]⁺

404.45 405 [M + 1]⁺

392.41 393 [M + 1]⁺

374.42 375 [M + 1]⁺

388.45 389 [M + 1]⁺

428.39 429 [M + 1]⁺

450.52 451 [M + 1]⁺

404.45 405 [M + 1]⁺

416.46 417 [M + 1]⁺

412.39 413 [M + 1]⁺

374.42 375 [M + 1]⁺

386.47 387 [M + 1]⁺

378.84 379 [M + 1]⁺

401.44 402 [M + 1]⁺

423.47 424 [M + 1]⁺

401.44 402 [M + 1]⁺

423.29 424 [M + 1]⁺

362.38 363 [M + 1]⁺

358.42 359 [M + 1]⁺

369.40 370 [M + 1]⁺

388.45 389 [M + 1]⁺

372.45 373 [M + 1]⁺

358.42 359 [M + 1]⁺

446.84 447 [M + 1]⁺

388.45 389 [M + 1]⁺

388.40 389 [M + 1]⁺

402.43 403 [M + 1]⁺

353.44 354 [M + 1]⁺

396.83 397 [M + 1]⁺

368.41 369 [M + 1]⁺

380.37 381 [M + 1]⁺ LCMS (ES) m/z, Structure MW [M + 1]+

376.44 377

367.47 368

365.46 366

365.46 366

407.54 408

379.48 380

337.40 338

405.52 406

359.41 360

351.43 352

351.43 352

310.38 311

451.54 452

379.48 380

351.43 352

367.47 368

379.48 380

379.48 380

379.48 380

326.38 327

312.35 313

350.32 351

364.35 365

399.47 400

Process 24

4-chlorothieno[3,2-c]quinoline (23 mg) was mixed with aniline (0.1 ml)and NMP (0.1 ml) and the mixture was heated in a microwave oven at 120°C. for 10 min. NMP (0.8 ml) was added and the compound purified bypreparative HPLC. Genevac evaporation affordedN-phenylthieno[3,2-c]quinolin-4-amine as a pinky solid (31 mg, quant.).LCMS (ES): 95% pure, m/z 277 [M+1]⁺.

Process 25

N1,N1-dimethyl-N2-(thieno[3,2-c]quinolin-4-yl)ethane-1,2-diamine wasprepared according to the procedure in process 24 using N,N-dimethylethylene diamine. Preparative HPLC and genevac evaporation afforded theexpected material as a TFA salt. LCMS (ES): 95% pure, m/z 272 [M+1]⁺.

Process 26

4-chlorothieno[3,2-c]quinoline-7-carboxylate (10 mg, 0.036 mmol) wassuspended in NMP (0.1 ml) and 3-aminomethylpyridine (0.1 ml). Themixture was heated in a microwave oven at 120° C. for 10 nm. Thereaction mixture was dissolved in a mixture of NMP and MeOH and theester intermediate purified by preparative HPLC. After genevacevaporation of the solvents, the resulting solid was dissolved in a 1:1mixture of THF and MeOH (0.6 ml). 5N aqueous LiOH (0.2 ml) was added andthe mixture stirred at room temperature for 17 hrs. Water and aqueousHCl were added and the solution of4-(pyridin-3-ylmethylamino)thieno[3,2-c]quinoline-7-carboxylic acid waspurified by preparative HPLC. Removal of the solvents by genevacevaporation provided compound4-(pyridin-3-ylmethylamino)thieno[3,2-c]quinoline-7-carboxylic acid as awhite solid (10 mg, 62% yield). LCMS (ES): 95% pure, m/z 336 [M+1]⁺. ¹HNMR (CDCl₃, 400 MHz) δ 5.23 (s, 2H), 7.71-7.78 (m, 4H), 8.11 (d, J=5.6,1H), 8.47 (d, J=8.0, 1H), 8.49 (d, J=0.8, 1H), 8.62 (d, J=5.2, 1H), 8.97(s, 1H) ppm.

Representative analogs (Table 6) were prepared by the same method, using4-chlorothieno[3,2-c]quinoline-7-carboxylate and appropriate amines. Thereaction temperatures used for the microwave reactions ranged from 120°C. to 180° C. After hydrolysis of the esters, the materials wereisolated by preparative HPLC/genevac evaporation. In some instances, thematerials precipitated after acidification of the hydrolysis mixture andwere isolated by filtration.

TABLE 6 LCMS Structure M.W. (ES) m/z

302.35 303 [M + 1]⁺

288.32 289 [M + 1]⁺

315.39 316 [M + 1]⁺

335.38 336 [M + 1]⁺

320.37 321 [M + 1]⁺

357.43 358 [M + 1]⁺

335.38 336 [M + 1]⁺

350.39 351 [M + 1]⁺

336.36 337 [M + 1]⁺

380.42 381 [M + 1]⁺

341.43 342 [M + 1]⁺

314.36 315 [M + 1]⁺

348.42 349 [M + 1]⁺

302.31 303 [M + 1]⁺

360.39 361 [M + 1]⁺

298.36 299 [M + 1]⁺

334.39 335 [M + 1]⁺

338.36 339 [M + 1]⁺

372.80 373 [M + 1]⁺

334.39 335 [M + 1]⁺

350.39 351 [M + 1]⁺

348.42 349 [M + 1]⁺

354.81 355 [M + 1]⁺

356.35 357 [M + 1]⁺

284.33 285 [M + 1]⁺

346.40 347 [M + 1]⁺

384.84 385 [M + 1]⁺

336.36 337 [M + 1]⁺

405.47 406 [M + 1]⁺

380.42 381 [M + 1]⁺

334.39 335 [M + 1]⁺

356.35 357 [M + 1]⁺

338.36 339 [M + 1]⁺

354.81 355 [M + 1]⁺

372.80 373 [M + 1]⁺

364.42 365 [M + 1]⁺

412.46 413 [M + 1]⁺

377.42 378 [M + 1]⁺

399.44 400 [M + 1]⁺

345.37 346 [M + 1]⁺

344.39 345 [M + 1]⁺

399.26 400 [M + 1]⁺

372.80 373 [M + 1]⁺

359.40 360 [M + 1]⁺

334.39 335 [M + 1]⁺

359.40 360 [M + 1]⁺

396.46 397 [M + 1]⁺

413.47 414 [M + 1]⁺

388.36 389 [M + 1]⁺

348.42 349 [M + 1]⁺

446.26 447 [M + 1]⁺

356.35 357 [M + 1]⁺

406.35 407 [M + 1]⁺

382.37 383 [M + 1]⁺

356.35 357 [M + 1]⁺

439.51 440 [M + 1]⁺

389.26 390 [M + 1]⁺

356.35 357 [M + 1]⁺

372.80 373 [M + 1]⁺

363.37 364 [M + 1]⁺ LCMS (ES) m/z Structure MW [M + 1]+

329.42 330

355.45 356

355.45 356

Process 27

4-(phenylamino)thieno[3,2-c]quinoline-7-carboxylic acid (6 mg) wasreacted with methyl sulfonamide (120 mg), EDCI (80 mg) and DMAP (20 mg)in anhydrous DMF (0.5 ml). After 5 hours, water was added and thesolution subjected to preparative HPLC. Genevac evaporation providedN-(methylsulfonyl)-4-(phenylamino)thieno[3,2-c]quinoline-7-carboxamideas a solid (6 mg, 81% yield). LCMS (ES): 95% pure, m/z 398 [M+1]⁺.

Process 28

In a vial, 4-oxo-4,5-dihydrothieno[3,2-c]quinoline-7-carboxylic acid(1.0 eq, 20 mg, 0.081 mmol), N-hydroxybenzotriazole monohydrate (2.0 eq,22 mg, 0162 mmol), para-methoxybenzylamine (2.0 eq, 21 ul, 0.162 mmol)and triethylamine (2.0 eq, 23 ul, 0.165 mmol) were dissolved inanhydrous DMF (0.5 ml). EDCI (2.0 eq 31 mg, 0.162 mmol) was added andthe reaction mixture was stirred at 70° C. overnight. MeOH (0.5 ml) andwater (2 ml) were added and the resulting precipitate filtered anddried. The material was triturated in AcOEt, filtered and dried toprovide an off-white solid (19 mg, 65% yield). LCMS (ES): 95% pure, m/z365 [M+1]⁺, ¹H NMR (DMSO-d₆, 400 MHz) δ 3.71 (s, 3H), 4.40 (d, J=6.0,2H), 6.88 (d, J=8.8, 2H), 7.24 (d, J=8.8, 2H), 7.60 (d, J=5.6, 1H), 7.69(dd, J=1.6, J=8.0, 1H), 7.84 (d, J=5.6, 1H), 7.90 (s, 1H), 7.91 (d,J=8.8, 1H), 9.11 (t, J=5.6, 1H) ppm

The following representative analogs (Table 7) were prepared by theseprocesses, using 4-oxo-4,5-dihydrothieno[3,2-c]quinoline-7-carboxylicacid and appropriate amines. In some instances, the materials werepurified by preparative HPLC and were isolated as dry solids afterGenevac evaporation.

TABLE 7 LCMS Structure M.W. (ES) m/z

315.39 316 [M + 1]⁺

372.44 373 [M + 1]⁺

320.37 321 [M + 1]⁺

316.33 316 [M + 1]⁺

327.38 328 [M + 1]⁺

380.42 381 [M + 1]⁺

405.47 406 [M + 1]⁺

321.35 322 [M + 1]⁺

350.39 351 [M + 1]⁺

354.81 355 [M + 1]⁺

338.38 339 [M + 1]⁺

357.43 358 [M + 1]⁺

314.36 315 [M + 1]⁺

286.35 287 [M + 1]⁺

349.41 350 [M + 1]⁺

302.35 303 [M + 1]⁺

408.47 409 [M + 1]⁺

272.32 273 [M + 1]⁺

355.41 356 [M + 1]⁺

284.33 285 [M + 1]⁺

334.39 335 [M + 1]⁺

378.40 379 [M + 1]⁺

413.49 414 [M + 1]⁺

427.52 428 [M + 1]⁺

364.42 365 [M + 1]⁺

339.37 340 [M + 1]⁺

335.38 336 [M + 1]⁺

348.42 349 [M + 1]⁺

335.38 336 [M + 1]⁺

335.38 336 [M + 1]⁺

467.56 468 [M + 1]⁺

The following representative analogs (Table 8) were prepared from theircorresponding methyl esters. The compounds were prepared according tothe hydrolysis procedure utilized for compound 15.

TABLE 8 Structure M.W. LCMS (ES) m/z

364.37 365 [M + 1]⁺

302.31 303 [M + 1]⁺

The following representative analogs (Table 9) were prepared from theircorresponding tert-butyl esters or N-Boc protected precursors. Theprecursors were treated with 30% trifluoroacetic acid in CH₂Cl₂ for 2hours. Removal of the volatiles in vacuo afforded the expectedmaterials.

TABLE 9 Structure M.W. LCMS (ES) m/z

327.40 328 [M + 1]⁺

313.37 314 [M + 1]⁺

316.33 317[M + 1]⁺

Process 29

ethyl 3-(7-(1H-tetrazol-5-yl)thieno[3,2-c]quinolin-4-ylamino)benzoate(1.0 eq, 7.6 mg, 0.018 mmol) was suspended in a 1:1:1 mixture of THF,MeOH and water. Lithium Hydroxide was added (40 mg, 1.66 mmol) and themixture stirred at room temperature for one hour. Water and hydrochloricacid were added and the resulting solid filtered and dried to afford3-(7-(1H-tetrazol-5-yl)thieno[3,2-c]quinolin-4-ylamino)benzoic acid as asolid. LCMS (ES): 95% pure, m/z 389 [M+1]⁺.

The following representative analogs (table 10) were prepared byreacting 3-(7-(1H-tetrazol-5-yl)thieno[3,2-c]quinolin-4-ylamino)benzoicacid and appropriate amines using the procedure described in process 28.The materials were purified by preparative HPLC and were isolated as drysolids after Genevac evaporation.

TABLE 10 LCMS (ES) Structure MW m/z

429.50 430[M + 1]⁺

457.51 458[M + 1]⁺

458.54 459[M + 1]⁺

459.48 460[M + 1]⁺

515.59 516[M + 1]⁺

478.53 479[M + 1]⁺

415.47 416[M + 1]⁺

427.48 428[M + 1]⁺

482.52 483[M + 1]⁺

445.50 446[M + 1]⁺

498.56 499[M + 1]⁺

Process 30

The following representative analogs (table 11) were prepared byreacting 3-(7-(methoxycarbonyl)thieno[3,2-c]quinolin-4-ylamino)benzoicacid and the appropriate amines using reaction conditions described inprocess 28. Hydrolysis of the ester using conditions described inprocess 29 afforded the following analogs.

TABLE 11 LCMS (ES) Structure MW m/z

405.47 406[M + 1]⁺

433.48 434[M + 1]⁺

439.49 440[M + 1]⁺

421.43 422[M + 1]⁺

434.51 435[M + 1]⁺

446.50 447[M + 1]⁺

491.56 492[M + 1]⁺

454.50 455[M + 1]⁺

391.44 392[M + 1]⁺

403.45 404[M + 1]⁺

458.49 459[M + 1]⁺

421.47 422[M + 1]⁺

474.53 475[M + 1]⁺

Process 31

The following representative analogs (table 12) were prepared byreacting2-(3-(7-(methoxycarbonyl)thieno[3,2-c]quinolin-4-ylamino)phenyl)aceticacid and the appropriate amines using reaction conditions described inprocess 30.

TABLE 12 LCMS (ES) Structure MW m/z

448.54 449[M + 1]⁺

417.48 418[M + 1]⁺

392.43 393[M + 1]⁺

405.47 406[M + 1]⁺

391.44 392[M + 1]⁺

Example 3 Processes for Synthesizing Compounds of Formulae IX, X, XI andXII Process 1

Methyl 2-amino-4-bromothiazole-4-carboxylate (1.0 eq, 100 mg, 0.42 mmol)was dissolved in anhydrous DMF (0.8 ml). The mixture was heated to 80°C. under nitrogen atmosphere. To the hot mixture, a solution oftent-Butyl nitrite (1.2 eq, 60 ul, 0.50 mmol) in DMF (0.8 ml) was addeddropwise. After a few minutes, absence of gas evolution indicatedcompletion of the reaction. The mixture was cooled down and poured ontoa prepacked silica gel column. Flash chromatography using hexanes, thenAcOEt/hexanes (2:8), provided methyl 5-bromothiazole-4-carboxylate as ayellow solid (49 mg, 53% yield). LCMS (ES): 95% pure, m/z 222 [M]⁺, 224[M+2]⁺.

Process 2

In a microwave vessel, methyl 5-bromothiazole-4-carboxylate (1.0 eq, 97mg, 0.44 mmol), 2-amino-3-methoxycarbonyl phenyl boronic acid HCl (1.1eq, 111 mg, 0.48 mmol), sodium acetate (3.0 eq, 107 mg, 1.31 mmol) andPdCl₂(dppf) (0.05 eq, 11 mg, 0.022 mmol) were mixed together inanhydrous DMF (1 ml). The mixture was heated in a microwave oven at 120°C. for 10 mn. Water was added and the material extracted with CH₂Cl₂.The combined extracts wre washed with brine, dried over Na₂SO₄ and thesolvents removed by evaporation. The material was dissolved in a mixtureof CH₂Cl₂ and MeOH and the solution filtered through a pad of celite.Evaporation of the volatiles afforded crude methyl4-oxo-4,5-dihydrothiazolo[4,5-c]quinoline-7-carboxylate as a black solid(44 mg, 39% yield). A small part of the compound was subjected topreparative HPLC for analytical purpose. LCMS (ES): 95% pure, m/z 261[M+1]⁺.

Process 3

Methyl 4-oxo-4,5-dihydrothiazolo[4,5-c]quinoline-7-carboxylate (35 mg,0.12 mmol) and LiOH (60 mg, 0.83 mmol) were stirred in a (1:1:1, v:v:v)mixture of THF, MeOH and water (0.6 ml) for 2 hours. 6N aqueous NaOH wasadded and the solution filtered through celite. The solution wasacidified and the resulting solid filtered. Preparative HPLCpurification and genevac evaporation provided4-oxo-4,5-dihydrothiazolo[4,5-c]quinoline-7-carboxylic acid as a whitesolid (0.8 mg). LCMS (ES): 95% pure, m/z 247 [M+1]⁺.

Process 4

Methyl 2-amino-4-bromothiazole-4-carboxylate (1.0 eq, 2.0 g, 8.44 mmol)was dissolved in CH₂Cl₂ (4 ml). Acetic anhydride (1.5 eq, 1.2 ml, 12.66mmol) and triethylamine (1.1 eq, 1.3 ml, 9.28 mmol) were added and themixture stirred at 100° C. for one hour. The resulting solid wasfiltered, triturated in AcOEt and then filtered again. After drying,methyl 2-acetamido-5-bromothiazole-4-carboxylate was isolated as a beigesolid (1.81 g, 77% yield). LCMS (ES): 95% pure, m/z 280 [M+1]⁺. ¹H NMR(CDCl₁, 400 MHz) δ 2.25 (s, 3H), 3.95 (s, 3H) ppm.

Process 5

Methyl2-acetamido-4-oxo-4,5-dihydrothiazolo[4,5-c]quinoline-7-carboxylate wasprepared according to the procedure used in process 2, starting frommethyl 2-acetamido-5-bromothiazole-4-carboxylate. Methyl2-acetamido-4-oxo-4,5-dihydrothiazolo[4,5-c]quinoline-7-carboxylate wasisolated as a black solid (106 mg, 37% yield). LCMS (ES): 95% pure, m/z318 [M+1]⁺.

Process 6

2-acetamido-4-oxo-4,5-dihydrothiazolo[4,5-c]quinoline-7-carboxylic acidwas prepared according to the procedure in process 3, starting from.Methyl2-acetamido-4-oxo-4,5-dihydrothiazolo[4,5-c]quinoline-7-carboxylate.-acetamido-4-oxo-4,5-dihydrothiazolo[4,5-c]quinoline-7-carboxylicacid was isolated as a black solid (14 mg, 44% yield). LCMS (ES): 95%pure, m/z 304 [M+1]⁺, ¹H NMR (DMSO-d₆, 400 MHz) δ 2.22 (s, 3H), 7.74(dd, J=1.2, J=8.0, 1H), 7.89 (d, J=8.4, 1H), 8.03 (d, J=1.6, 1H), 12.07(s, 1H), 12.80 (s, 1H) ppm.

Process 7

2-acetamido-4-oxo-4,5-dihydrothiazolo[4,5-c]quinoline-7-carboxylic acid(102 mg, 0.34 mmol) was stirred at 120° C. in aqueous 6N HCl overnight.Water was added and the compound was filtered and dried to provide2-amino-4-oxo-4,5-dihydrothiazolo[4,5-c]quinoline-7-carboxylic acid as ablack solid (76 mg, 86% yield). LCMS (ES): 95% pure, m/z 262 [M+1]⁺, ¹HNMR (DMSO-d₆, 400 MHz) δ 7.60 (d, J=8.4, 1H), 7.70 (dd, J=1.2, J=8.0,1H), 7.99 (d, J=1.2, 1H), 11.94 (s, 1H) ppm.

Process 8

Methyl 4-oxo-4,5-dihydrothiazolo[4,5-c]quinoline-7-carboxylate (1.0 eq,0.62 g, 2.38 mmol) was suspended in toluene. DIEA (1.5 eq, 122 ul, 3.57mmol) and POCl₃ (2.3 eq, 507 ul, 5.47 mmol) were added and the mixturevigorously stirred at 120° C. for 1 hour. Water, ice and CH₂Cl₂ wereadded and the resulting emulsion filtered through celite. The organicphase was decanted and the aqueous phase further extracted with CH₂Cl₂.The combined organic extracts were dried over Na₂SO₄ and the solventremoved in vacuo to afford methyl4-chlorothiazolo[4,5-c]quinoline-7-carboxylate (0.31 g, 47% yield). LCMS(ES): >90% pure, m/z 279[M+1]⁺.

Process 9

In a microwave vessel, methyl4-chlorothiazolo[4,5-c]quinoline-7-carboxylate (1.0 eq, 23 mg, 0.084mmol) and aniline (13 eq, 0.1 ml, 1.1 mmol) were mixed in NMP (0.1 ml).The mixture was heated in a microwave oven at 120° C. for 10 min. Theintermediate ester was purified by preparative HPLC and isolated as asolid after genevac evaporation. The solid was stirred in a (1:1:1,v:v:v) mixture of THF, MeOH and water (0.6 ml) with LiOH (41 mg) at roomtemperature for 2 hours. HCl and water were added, the organic solventswere evaporated and the solution allowed resting for 2 hours. Theprecipitate that slowly formed was filtered and dried to afford4-(phenylamino)thiazolo[4,5-c]quinoline-7-carboxylic acid as a solid (8%yield over 2 steps). LCMS (ES): >95% pure, m/z 322 [M+1]⁺.

Representative analogs (Table 13) were prepared by the same processusing methyl 4-chlorothiazolo[4,5-c]quinoline-7-carboxylate andappropriate amines. The reaction temperatures used for the microwavereactions ranged from 120° C. to 180° C. After synthesis of the finalcompounds, the materials were isolated by preparative HPLC/genevacevaporation. In some instances, the materials precipitated afteracidification and were isolated by filtration.

TABLE 13 LCMS (ES) Structure MW m/z

345.37 346 [M + 1]⁺

339.34 340 [M + 1]+

373.79 374 [M + 1]+

351.38 352 [M + 1]+

Example 4 Modulation of CK2 and PARP Activity in Cell-Free In VitroAssays

Modulatory activity of compounds described herein was assessed in vitroin cell-free CK2 assays. Modulatory activity of compounds describedherein also are assessed in vitro in cell-free PARP assays. These assaysare described hereafter.

CK2 Assay

Test compounds in aqueous solution were added at a volume of 10microliters, to a reaction mixture comprising 10 microliters AssayDilution Buffer (ADB; 20 mM MOPS, pH 7.2, 25 mM beta-glycerolphosphate,5 mM EGTA, 1 mM sodium orthovanadate and 1 mM dithiothreitol), 10microliters of substrate peptide (RRRDDDSDDD, dissolved in ADB at aconcentration of 1 mM), 10 microliters of recombinant human CK2 (25 ngdissolved in ADB; Upstate). Reactions were initiated by the addition of10 microliters of ATP Solution (90% 75 mM MgCl₂, 75 micromolar ATPdissolved in ADB; 10% [γ-³³P]ATP (stock 1 mCi/100 μl; 3000 Ci/mmol(Perkin Elmer) and maintained for 10 minutes at 30 degrees C. Thereactions were quenched with 100 microliters of 0.75% phosphoric acid,then transferred to and filtered through a phosphocellulose filter plate(Millipore). After washing each well 5 times with 0.75% phosphoric acid,the plate was dried under vacuum for 5 min and, following the additionof 15 ul of scintilation fluid to each well, the residual radioactivitywas measured using a Wallac luminescence counter.

PARP Assay

PARP assays are conducted using a chemiluminescent PARP assay kit(Trevigen). Briefly, reactions are performed in Histone-coated stripwells, by adding 10 microliters test compound dissolved in 1×PARP Buffer(prepared by mixing 20×PARP buffer diluted with high-purity water) and15 microliters diluted PARP-HSA enzyme (diluted in 1×PARP buffer, 0.1unit per well) to 25 microliters PARP cocktail (prepared from 10× PARPcocktail and 10× activated DNA, both 2.5 microliters per well and 20microliters per well of 1×PARP buffer). The reactions are incubated atambient temperature for 60 minutes, then the liquid was removed. Afterwashing the wells four times with PBS (200 ul), 50 microliters ofSTREP-HRP (Horseradish Peroxidase) solution (diluted 500-fold in 1×Strep-Diluent) was added and the reactions were allowed to incubate for30 minutes at ambient temperature. The liquid was removed and, afterwashing the wells four times with PBS (200 ul), 50 microliters each ofPeroxyGlo A and B (Chemiluminescent Horseradish Peroxidase substrates)are added and the resulting chemiluminescence quantified on theSpectraMax M5 plate reader.

Tables 14A, 14B, and 15-18 show modulatory effects of compounds on CK2and/or PARP activity.

TABLE 14A CK2 PARP Compound Inhibition Inhibition

28% (at 5 μM) IC₅₀ = 0.070 μM

29% (at 5 μM) IC₅₀ = 0.060 μM

38% (at 5 μM) IC₅₀ = 0.40 μM

IC₅₀ = 2 μM IC₅₀ = 0.030 μM

IC₅₀ = 0.18 μM IC₅₀ = 1.0 μM

IC₅₀ = 2.5 μM IC₅₀ = 0.80 μM

IC₅₀ = 1.0 μM 15% (at 1 μM)

IC₅₀ = 1.6 μM 9% (at 1 μM)

16% (at 2.5 μM) 33% (at 1 μM)

IC₅₀ = 0.013 μM

96% (at 1 μM)

46% (at 1 μM)

78% (at 1 μM)

62% (at 1 μM)

TABLE 14B CK2 IC50 CK2 % inhibition Structure (uM) 5 uM 2.5 uM 1.0 uM

1.2

>10

>10

0.67

1.1

0.27

0.95

0.32

0.9

1.22

0.43

0.55

0.35

2

84%

>5

63%

 0%

 0%

28%

78%

 0%

 0%

29%

0.19

1.5

0.31

0.15

1.1

0.12

18%

0.21

0.67

0.97

0.58

0.43

0.82

1.17

0.43

 5%

 0%

 0%

70%

 0%

 0%

 0%

 0%

71%

84%

80%

77%

75%

61%

65%

68%

77%

60%

Table 15 shows modulatory effects of compounds on PARP and CK2.

TABLE 15 CK2 PARP PARP % inhib CK2 % inhib @ % inhib PARP @ 10 IC50Structure 20 uM @ 1 uM IC50(uM) uM (uM)

0 • • 0 •

85 • • • •

90 58 1 77 4

84 27 • 17

84 39 • 5

82 40 • 8 •

22 0 • 22 •

93 47 • 10 •

95 35 • 16

97 31 • 12

52 0 • 10

32 0 • 3

37 0 • −3

62 0 • −9

24 0 • −7

55 0 • −10

97 83 0.2 7

96 77 0.5 −9

95 82 0.4 2

88 65 1 −34

83 55 1 −24

93 65 0.4 −19

67 15 • −22

97 89 0.2 3

94 71 0.3 7 •

90 69 0.5 0 •

• 36 • 14 •

• • • −1 •

• 24 • 5 •

• • • −16 •

• 72 0.3 −25 •

• 49 • 10 •

• • • 1 •

• 27 • 8 •

• 67 0.5 −13 •

• 45 • 1 •

• 71 1 3 •

• 64 0.5 1 •

• 75 1 −13 •

• 71 • −24 •

• 29 • −1 •

• 96 0.03 −27 •

• 96 0.02 −3 •

• 12 • 41 •

• 79 0.06 −14 •

• 74 0.4 3 •

• 21 • 48 2.8

• 51 0.5 −5 •

• 39 • 86 0.9

• 5 • 44 12.5

• 18 • 18 •

• 40 •

TABLE 16 CK2: IC50 CK2: IC50 (uM) (uM) Structure (15 uM ATP) (20 um ATP)

0.006 0.01

0.025 0.019

0.07 0.06

0.311 0.13

0.113 0.2

0.004 0.007

0.004 0.006

1.469 1.661

25

0.01

0.005

0.003

0.002

0.651

0.006

0.006

0.007

0.006

0.047

0.052

0.019

0.007

0.003

0.045

0.009

0.005

0.007

0.016

0.005

0.004

>0.5

>0.5

>0.5

>0.5

0.711

0.018

0.027

0.051

0.069

0.02

0.026

0.056

0.163

0.107

0.089

0.046

0.06

0.04

0.144

0.25

0.009

0.018

0.013

0.011

>0.75

0.018

>0.75

0.004

0.134

0.009

0.03

0.02

0.007

0.083

0.052

0.171

0.107

0.349

0.114

0.05

0.214

0.172

>0.75

>0.75

>0.75

0.028

0.021

>0.75

0.493

0.006

0.059

0.026

>0.75

0.006

0.011

0.102

0.086

0.134

0.018

0.035

>0.75

0.168

0.686

0.356

0.103

>0.75

>0.75

>0.75

0.513

0.027

0.185

0.016

>0.75

>0.75

>0.75

0.023

0.015

0.014

>0.75

0.087

>0.75

0.014

0.093

0.01

0.035

0.033

0.02

0.198

TABLE 17 CK2: IC50 CK2: (uM) IC50(uM) Structure (15 uMATP) (20 uM ATP)

0.995 1.2

0.748 0.67

1.258 1.1

0.102 0.277

0.622 0.872

0.092 0.31

0.367 0.9

0.922 1.22

0.168 0.518

0.171 0.55

0.507 0.369

0.771 2

0.231 0.28

0.516 1.006

0.096 0.189

1.5

0.219 0.31

0.15

1.1

0.12

0.21

0.67

0.97

0.32 0.58

0.131 0.43

0.257 0.82

0.666 1.17

0.238 0.431

0.252 0.31

0.371 0.372

0.194 0.382

0.172 0.3

0.233 0.407

0.256 0.462

0.358 10

0.611 0.392

0.42 0.27

0.348 0.35

0.812 0.89

0.458 0.406

0.154 0.216

0.129 0.181

0.171 0.283

0.198 0.268

0.485 0.524

0.122 0.14

0.075 0.096

0.235 0.375

0.346 0.423

0.358 0.509

0.29 0.63

0.135

0.07

0.068

0.032

0.07

0.126

0.395

0.129

0.103

0.081

0.028

0.38

0.502

0.549

0.24

0.363

0.318

0.237

0.288

0.251

0.303

0.224

0.307

0.192

0.366

0.221

0.137

0.187

0.335

0.156

0.09

0.121

0.281

0.061

0.242

0.091

0.256

0.156

0.127

0.138

0.116

0.035

0.127

0.076

0.131

0.289

0.141

0.204

TABLE 18 CK2: IC50 (uM) CK2: IC50 (uM) Structure (15 uM ATP) (20 um ATP)

4.7

3.4

0.169 0.219

0.037

0.12

0.146

0.044

Example 5 Cell Proliferation Modulatory Activity

A representative cell-proliferation assay protocol using Alamar Blue dye(stored at 4° C., use 20 ul per well) is described hereafter.

96-Well Plate Setup and Compound Treatment

-   -   a. Split and trypsinize cells.    -   b. Count cells using hemocytometer.    -   c. Plate 4,000-5,000 cells per well in 100 μl of medium and seed        into a 96-well plate according to the following plate layout.        Add cell culture medium only to wells B10 to B12. Wells B1 to B9        have cells but no compound added.

1 2 4 5 7 8 10 11 3 6 9 12 A EMPTY B NO COMPOUND ADDED Medium Only C 10nM 100 nM 1 uM 10 uM Control D 10 nM 100 nM 1 uM 10 uM Comp1 E 10 nM 100nM 1 uM 10 uM Comp2 F 10 nM 100 nM 1 uM 10 uM Comp3 G 10 nM 100 nM 1 uM10 uM Comp4 H EMPTY

-   -   d. Add 100 μl of 2× drug dilution to each well in a        concentration shown in the plate layout above. At the same time,        add 100 μl of media into the control wells (wells B10 to B12).        Total volume is 200 μl/well.    -   e. Incubate four (4) days at 37° C., 5% CO2 in a humidified        incubator.    -   f. Add 20 μl Alamar Blue reagent to each well.    -   g. Incubate for four (4) hours at 37° C., 5% CO2 in a humidified        incubator.    -   h. Record fluorescence at an excitation wavelength of 544 nm and        emission wavelength of 590 nm using a microplate reader.

In the assays, cells are cultured with a test compound for approximatelyfour days, the dye then is added to the cells and fluorescence ofnon-reduced dye is detected after approximately four hours. Differenttypes of cells can be utilized in the assays (e.g., HCT-116 humancolorectal carcinoma cells, PC-3 human prostatic cancer cells andMiaPaca human pancreatic carcinoma cells). Anti-proliferative effects ofrepresentative compounds are provided hereafter

TABLE 19A IC50 IC50 IC50 IC50 IC50 IC50 (uM) (uM) (uM) IC50 (uM) (uM)(uM) (uM) Structure A549 MCF-7 LNCaP MDAMB231 Raji HL-60 K-562

4.16 10.79 8.18 2.66 13.70 4.86 4.01

6.83 8.24 4.57 6.13 4.51 1.92 4.95

1.11

16.65

47.04 14.71 8.60

6.59 17.68 4.89 6.66 3.32 2.64 2.99

24.58 2.02 1.83 3.10 8.47 1.85 2.41

14.10 1.06 1.36 0.84 4.51 9.68 1.77

28.46 1.79 1.56 1.18 7.35 1.13

21.21 1.27 1.40 4.25 3.38 4.49 1.20

>50 >50 <0.2 >50 40.62

>50 5.94 48.24 >50 >50

13.86 3.40 1.44 2.38 4.97 0.73 1.68

9.74 0.76 7.39 3.79 5.46 3.74 8.65

30.24 1.43 17.08 11.80 4.28 5.59 3.33

>50 >50 37.38 >50 31.21

37.98

32.50 47.63 13.91 14.22 9.18

47.17 >50 10.30 5.83 8.11

>50 >50 10.43 7.66 7.17

27.37 1.89 10.76 11.04 6.35 4.81 3.26

>50 40.95 15.51 28.65 9.15

0.73

18.16

24.45

>50

48.21

>50

10.51

2.44

>50

4.90

10.44

4.74

>50

12.45

5.21

4.43

3.93

2.93

26.52

8.28

9.82

4.12

20.77

9.19

6.87

15.77

6.53

7.12

12.63

31.58

5.22

7.05

8.38

2.63

>50

5.48

>50

27.18

7.23

>50

>50

>50

>50

7.22

23.54

6.88

>50

17.50

13.02

23.04

12.77

20.11

>50

>50

>50

>50

9.66

33.72

25.43

>50

39.84

10.47

>50

5.48

12.11

19.23

>50

4.27

34.23

>50

2.52

>50

4.36

Example 6 Modulation of Endogenous CK2 Activity

The human leukemia Jurkat T-cell line was maintained in RPMI 1640(Cambrex) supplemented with 10% fetal calf serum and 50 ng/mlGeutamycin. Before treatment cells were washed, resuspended at a densityof about 10⁶ cells/milliliter in medium containing 1% fetal calf serumand incubated in the presence of indicated mounts of drug for two hours.Cells were recovered by centrifugation, lysed using a hypotonic buffer(20 mM Tris/HCl pH 7.4; 2 mNI EDTA; 5 mM EGTA; 10 mM mercaptoethanol; 10mM NaF; 1 uM Okadaic acid; 10% v/v glycerol; 0.05% NP-40; 1% ProteaseInhibitor Cocktail) and protein from the cleared lysate was diluted to 1microgram per microliter in Assay Dilution Buffer (ADB; 20 mM MOPS, pH7.2, 25mM β-glycerolphosphate, 5 mM EGTA, 1mM sodium orthovanadate and1mM dithiothreitol). To 20 microliters of diluted protein was added 10microliters of substrate peptide (RRRDDDSDDD, dissolved in ADB at aconcentration of 1 mM) and 10 microliters of PKA Inhibitor cocktail(Upstate). Reactions were initiated by the addition of 10 microliters ofATP Solution (90% 75 mM MgCl₂, 100 uM ATP dissolved in ADB; 10%[gamma-³³P]ATP (stock 1 mCi/100 microliters; 3000Ci/mmol (Perkin Elmer))and maintained for 15 min at 32 degrees C. The reactions were quenchedwith 100 microliters of 0.75% phosphoric acid, then transferred to andfiltered through a phosphocellulose filter plate (Millipore). Afterwashing each well 5 times with 0.75% phosphoric acid, the residualradioactivity was measured using a Wallac luminescence counter.

Modulatory activities of two compounds assessed by the assay are shownin FIG. 1. Structures of the compounds are provided below:

As shown in FIG. 1, each of the two compounds significantly inhibitedendogenous CK2 activity as compared to the untreated control. Each ofthe two compounds also more potently inhibited endogenous CK2 activityas compared to reference compound 4,5,6,7-tetrabromobenzotriazole (TBB),a known CK2 inhibitor (Ruzzene et al., Biochem J. 15: 364(Pt 1):41-7(2002)).

TABLE 20 Modulation of endogenous CK2 activity Modulation of endogenousCK2 activity Structure IC50 (uM)

25.8

4.338

3.564

10.66

8.36

50

15.7

50

9.59

37.89

4.426

0.58

TABLE 21 Modulation of endogenous CK2 activity Structure Modulation ofendogenous CK2 activity IC50 (uM

7.4

>50

19.87

2.325

0.464

7.066

>50

>50

1.056

2.933

0.688

0.1

0.269

0.026

0.098

0.63

0.22

0.017

0.07

1.016

0.64

3.6

2.5

1.351

0.01

0.01

0.098

0.044

0.01

0.01

0.044

0.03

0.047

0.172

0.011

0.027

Example 7 Evaluation of Pharmacokinetic Properties

The pharmacokinetics properties of drugs were investigated in ICR micefollowing an intravenous (IV) bolus and oral (PO) doses of drug at 5mg/kg and 25 mg/kg respectively. Blood samples were collected atpredetermined times and the plasma separated. Plasma was separated fromthe blood samples collected at 5, 15 and 30 minutes and 1, 2, 4, 8 and24 hours post-dose.

The pharmacokinetics properties of drugs were also investigated in SDrats and beagle dogs following an intravenous (IV) bolus and oral (PO)doses of drug using similar methods. Blood samples were collected atpredetermined times and the plasma separated.

Drug levels were quantified by the LC/MS/MS method described below.Noncompartmental pharmacokinetic analysis was applied for intravenousadministration. A linear trapezoidal rule was used to compute AUC(0-24).The terminal t_(1/2) and C₀ were calculated using the last three and thefirst three data points, respectively

Bioanalysis was performed using a Quattro Micro LC/MS/MS instrument inthe MRM detection mode, with an internal standard (1S). Briefly, 15 □Lplasma samples were prepared for analysis using protein precipitationwith 120 μL of acetonitrile. The supernatants were transferred into a 96well plate and subjected to LC-MS/MS analysis using a PhenomenexPolar-RP HPLC column. The mobile phases were 10 mM NH₄HCO₃ in water(Solution-A) and 10 mM NH₄HCO₃ in methanol (Solution-B). The column wasinitially equilibrated with 25% Solution-B and followed with 100%Solution B over 5 minutes. The method had a dynamic range from 1 to10,000 ng/mL. Quantitation of the analytes was performed in the batchmode with two bracketing calibration curves according to thebioanalytical sample list.

Pharmacokinetic profiles and estimated pharmacokinetic parameters ofcompound A1 below are shown in FIG. 2A and in Table 22.

TABLE 22 Estimated pharmacokinetic parameters after intravenous and oraldosing at 5 and 25 mg/kg, respectively in ICR mice. PK Parameter IV POUnits Dose 5 25 mg/kg AUC_((0-8 h)) 2910 1580 AUC_((0-24 h)) 3337 2915ng · h · ml⁻¹ AUC_((0-Inf)) 3364 3149 ng · h · ml⁻¹ Cmax-obs N/A 343ng/mL Cp0-exp 13201 N/A ng/mL Tmax N/A 0.25 hr Kel 0.1586 0.1076 hr⁻¹t_(1/2) 4.4 6.4 hr Vd 9.4 N/A L/kg CL_(s) 1.5 N/A L/kg/hr F(0-8 h) N/A10.9 % F(0-inf h) N/A 18.7 %

Pharmacokinetic profiles and estimated pharmacokinetic parameters of thetest compound below are shown in FIG. 2B and Table 23.

TABLE 23 Estimated pharmacokinetic parameters after IV and PO dose inICR mice. PK Parameter IV PO Unit Dose 3.4 24.5 mg/kg AUC_((0-8 h)) 37166005 AUC_((0-24 h)) 4806 9120 ng · h · ml⁻¹ AUC_((0-Inf)) 4898 10895 ng· h · ml⁻¹ Cmax-obs 4744 1600.5 ng/mL Cp0-exp 5631 N/A ng/mL Tmax N/A0.5 hr Kel 0.1418 0.0594 hr⁻¹ t_(1/2) 4.9 11.7 hr Vd 4.9 N/A L/kg CL_(s)0.7 N/A L/kg/hr F_((0-24 h)) N/A 26.5 % F_((0-Inf)) N/A 31.1 %

Pharmacokinetic profiles and estimated pharmacokinetic parameters of thetest compound A1 in dogs are shown in Table 24. Pharmacokinetic profilesand estimated pharmacokinetic parameters of the test compound A1 in ratsare shown in Table 25.

TABLE 24 Estimated pharmacokinetic parameters of test compound A1 afterIV and PO dose in Beagle dogs. PK Parameters IV PO IV MC PO MC Unit Dose0.80 3.80 0.80 3.80 mg/kg AUC_((0-8 h)) 345 1024 633 1775 AUC_((0-12 h))349 1064 N/A N/A ng · h · mI⁻¹ AUC_((0-Inf)) 352 1073 633 1804 ng · h ·mI⁻¹ Cmax-obs 1043 494.7 1979 908 ng/mL Cp0-exp 1406 N/A 2723 N/A ng/mLTmax N/A 0.5 N/A 0.25 hr Kel 0.5546 0.5546 N/D 0.4318 hr⁻¹ t_(1/2) 1.21.2 N/D N/D hr Vd 4.1 N/A N/D N/D L/kg CL_(s) 2.3 N/A N/D N/D L/kg/hrF_((0-8 h)) N/A 62.5 N/A 59.0 % F_((0-12 h)) N/A 64.2 N/A N/A %F_((0-Inf)) N/A 64.1 N/A N/A %

TABLE 25 Estimated pharmacokinetic parameters of test compound A1 afterIV and PO dose in SD rats. PK Parameter IV PO Unit Dose 2.50 12.50 mg/kgAUC_((0-12 h)) 13119 19025 ng · h · mI⁻¹ AUC_((0-24 h)) 14352 25858 ng ·h · mI⁻¹ AUC_((0-Inf)) 13997 26587 ng · h · mI⁻¹ Cmax-obs 21339 4207.4ng/mL Cp0-exp 27117 N/A mg/mL Tmax N/A 1.0 hr Kel 0.0707 0.1529 hr⁻¹t_(1/2) 9.8 4.5 hr Vd 2.5 N/A L/kg CL_(s) 0.2 N/A L/kg/hr F_((0-12 h))N/A 29.0 % F_((0-24 h)) N/A 36.0 % F_((0-Inf)) N/A 38.0 %

Pharmacokinetic profiles and estimated pharmacokinetic parameters of thetest compound A2 in beagle dogs and SD rats are shown in Table 26.

TABLE 26 Estimated pharmacokinetic parameters of test compound A2 afterIV and PO dose. PK Parameter IV SD Rats PO SD-Rats IV Dog PO Dog UnitDose 1.56 8.06 2.00 7.50 mg/kg AUC_((0-8 h)) 35755 55808 1394 2253AUC_((0-12 h)) 39194 73945 1414 2315 ng · h · mI⁻¹ AUC_((0-Inf)) 3665980286 1437 2355 ng · h · mI⁻¹ Cmax-obs 34264.8 6668.3 3070.8 1212.8ng/mL Cp0-exp 47935 N/A 3847 N/A ng/mL Tmax N/A 2.0 N/A 1.0 hr Kel0.1215 0.1077 0.1360 0.2092 hr⁻¹ t_(1/2) 5.7 6.4 5.1 3.3 hr Vd 0.6 N/A10.2 N/A L/kg CL_(s) 0.1 N/A 1.4 N/A L/kg/hr F_((0-8 h)) N/A 31.2 N/A43.1 % F_((0-12 h)) N/A 37.7 N/A 43.7 % F_((0-Inf)) N/A 43.8 N/A 43.7 %

Example 8 Evaluation of Compound Efficacy in Tumor Suppression

The in vivo activity of compound A1 and compound A2 (shown previously)was assessed by intravenous and oral administration to tumor-bearingxenograft mice. The in vivo experiments followed protocols approved bythe Animal Use and Care Committee. Female NCr nu/nu mice were purchasedfrom Taconic Farms and group housed in a ventilated rack system on a12/12 light cycle. All housing materials and water were autoclaved priorto use. The mice were fed ad libitum with ganuna irradiated laboratorychow and acidified water. Animals were handled under laminar-flow hoods.

Tumor size (mm³) was calculated using the formula (l×w²)/2, wherew=width and l=length in mm of the tumor. Tumor weight was estimated withthe assumption that 1 mg is equivalent to 1 mm³ of tumor volume.

For intravenous administration of compound A1, animals were inoculatedsubcutaneously in the right flank with 5×10⁶ MiaPaca cells. Tumors weremonitored twice weekly and then daily as they approached the appropriatesize for study. On Day 1 of the study, the animals were randomized inton=5 treatment groups with group mean tumor sizes of 160 mm³.

Grp 1 Mean 160.966 UTC Grp 2 Mean 161.816 Gemzar Grp 3 Mean 161.807 30mg/kg CK2 Compound Grp 4 Mean 159.621 60 mg/kg CK2 Compound % Dif. 1.363SD 1.034.

Animals received 14 doses of Vehicle, Gemzar at 100 mg/kg Q3D orcompound A1 at either 30 mg/kg or 60 mg/kg by QD intravenousadministration. Tumor volume measurements (FIG. 3A) and body weight(FIG. 3B) were recorded on days 3, 6, 8, 10, 13 and 15. Photographs ofspecific untreated control animals and animals administered 60 mg/kgcompound A1 are shown in FIGS. 3C and 3D. Compound A1 is referred to as“CK2 inhibitor” in FIGS. 3A, 3B, 3C and 3D.

Compound A1 also was administered orally to MiaPaca xenograft animalsand inhibited tumor growth. Compound A1 was formulated as a sodium saltat 10 mg/mL with 2% PEG 300 and buffered to pH 8.4 using sodiumphosphate buffer. Compound A1 when administered orally to the animals ata dose of 100 mg/kg QD×8 and then 200 mg/kg QD×5 significantly inhibitedtumor growth relative to an untreated control group. Gemzar™administered at a dose of 80 mg/kg IP Q3D was used as a positivecontrol. Compound A1 also was delivered by oral administration at 100mg/kg to animals bearing MCF-7 xenografts and at 150 mg/kg to animalsbearing PC-3 xenografts, and in both sets of studies, significantlyinhibited tumor growth.

It also was determined that compound A1 reduced CK2 activity in tumors.Assessment of CK2 activity in tumors revealed that tumors from animalstreated with compound A1 had about 40% of the CK2 activity of tumorsfrom animals not treated with compound A1 or treated with Genizar™.

The distribution of compound A1 in the plasma and tumors of animals wasassessed. In animals administered 30 mg/kg compound A1 IV, 60 mg/kgcompound A1 IV and 200 mg/kg compound A1 orally, about 6.8, 2.2 and 9.5micromolar compound A1, respectively, was identified in plasma, andabout 42.9, 7.0 and 6.4 micromolar compound A1, respectively, wasidentified in tumors.

Caspase staining also was assessed as a biomarker for compound A1treatment of tumors. In animals treated with 60 mg/kg of compound A1 byIV administration, caspase-3 cell staining levels were four-fold greaterthan in untreated control cells. These results suggest caspase-3staining can be a useful biomarker for monitoring inhibition of cellproliferation and tumor inhibition.

It was also determined that compound A1 significantly inhibited tumorgrowth in A549 (human lung cancer cells) and BX-PC3 (human pancreaticcancer cells) xenograft mice. The compound was delivered by oraladministration for such determinations.

For assessment of compound A2, the compound was delivered by intravenousand intraperitoneal administration to tumor-bearing xenograft mice.Animals were inoculated subcutaneously in the right flank with 5×10⁶BC-PC3 cells. Tumors were monitored twice weekly and then daily as theyapproached the appropriate size for study. On Day 1 of the study, theanimals were randomized into n=8 treatment groups (n=5 for positive andnegative control groups) with group mean tumor sizes of 97 mm³.

Grp 1 Mean 97.80 UTC Grp 2 Mean 96.95 Gemzar Q3D Grp 3 Mean 96.68 50mg/kg CX-5011 IV BID × 10 days Grp 4 Mean 98.95 60 mg/kg CX-5011 IV QD ×17 days Grp 5 Mean 96.51 100 mg/kg CX-5011 IP BID × 17 days % Dif 2.50SD 1.01

Animals received 17 doses of Vehicle, Gemzar at 100 mg/kg Q3D orcompound at either 60 mg/kg QD intravenous administration or 100 mg/kgBID intraperitoneal administration. One group (#3) received 10 doses ofcompound at 50 mg/kg BID intravenous administration. Tumor volumemeasurements and body weight were recorded on days 1, 4, 7, 11, 13, 15,and 18, and data showed compound A2 significantly inhibited tumorprogression (FIG. 4A) while not significantly altering body weight (FIG.4B). Delivery of compound A2 to animals bearing MiaPaca xenografts by IVadministration at 50 and 60 mg/kg and by IP administration at 100 mg/kgsignificantly inhibited tumor progression. Also, delivery of compound A2to animals bearing MDA-MB-231 xenografts by IV administration at 30 and60 mg/kg and by oral administration at 200 mg/kg significantly inhibitedtumor progression. Delivery of compound A2 to animals bearing MiaPacaxenografts by oral administration at 100 mg/kg QD×8 and 200 mg/kg QD×6significantly inhibited tumor progression. A meglumine salt of compoundA2 at pH 10.0 and at 10 mg/mL was utilized as an oral formulation forthe studies.

Tumor pharmacokinetic studies of compound A2 were carried out in which30 mg/kg of the compound was dosed IV QD×6. Plasma, blood and tumorsamples were taken on day 1, 4 and 6 and three animals sacrificed foreach time point. Steady state was reached after about three days, theterminal slope decreases, the half life about doubles, the minimumconcentration was 4-5 times higher after six days and there were nosignificant differences between day 4 and 6.

Delivery of compound A3 to animals bearing MiaPaca xenografts by IVadministration also significantly inhibited tumor progression.

Example 9 Modulation of Non-CK2 Protein Kinase Activity

Compounds described herein are profiled for in vitro modulatory activityagainst protein kinases other than CK2. The in vitro analysis isconducted using known protocols (e.g., assay protocols described atworld-wide web address upstate.com/img/pdf/KP_AssayProtocol_Booklet_v3.pdf). Compounds described herein are screened in theassays and prioritized based upon modulatory activity against proteinkinases other than CK2 and specificity for CK2 or PARP.

Example 10 Evaluation of Angiogenesis Inhibition by Endothelial TubeFormation Assay

A human endothelial tube formation assay was performed using the 96-wellBD BioCoat™ Angiogenesis System from BD Biosciences, using themanufacturer's recommended protocol.

Briefly, HUVEC cells (from ATCC) were suspended in 150 ul of mediacontaining 10% FBS at 4×10⁵ cells/ml in each of the 96-wells of thematrigel coated plate in the presence or absence of variousconcentrations of compound A2. The plate was incubated for 18 hrs at 37°C. The cells were stained with calcein AM and the results visualized byfluorescent microscopy or by phase contrast. It was observed thatcompound A2 inhibited tube formation in the assay described above over aconcentration range of 1 to 5 μM.

Example 11 Modulation of Protein Kinase Activity in Cell-Free In VitroAssay

The biological activity of several compounds were tested in variousprotein kinase assays.

Modulation of PIM-1 Kinase Activity in Cell-Free In Vitro Assay

Test compounds (10 ml) dissolved in 95% 20 mM MOPS pH7.2, 5% DMSO wereadded to a reaction mixture comprising 10 ul of 5× Reaction Buffer (40mM MOPS pH 7.0, 5 mM EDTA), 10 ul of substrate peptide (KKRNRTLTV,dissolved in water at a concentration of 1 mM), 10 ml of recombinanthuman PIM1, 4 ng dissolved in PIM1 dilution buffer (20 mM MOPS pH 7.0;EDTA 1 mM; 5% Glycerol; 0.01% Brij 35; 0.1%; 0.1% 2-mercaptoethanol; 1mg/ml BSA). Reactions were initiated by the addition of 10 ul of ATPSolution (49% (15 mM MgCl₂; 75 uM ATP) 1% ([γ-33P]ATP: Stock 1mCi/100μl; 3000Ci/mmol (Perkin Elmer)) and maintained for 10 min at 30° C. Thereactions were quenched with 100 ul of 0.75% Phosphoric acid, thentransferred to and filtrered through a Phosphocellulose filter plate(Millipore). After washing each well 4 times with 0.75% Phosphoric acid,the residual radioactivity was measured using a Wallac luminescencecounter.

Modulation of FLT-3 Kinase Activity in Cell Free In Vitro Assay

FLT-3 Inhibition was determined by measuring the inhibition ofrecombinant human FLT-3 phosphorylation of the peptide EAIYAAPFAKKKusing 10 uM ATP in a reaction mixture containing 20 mM Hepes pH 7.5, 10mM MgCl₂, 1 mM EGTA, 0.02% Brij35, 0.02 mg/ml BSA, 0.1 mM Na₃VO₄, 2 mMDTT, and 1% DMSO.

Modulation of Protein Kinase Activity in Standardized Radiometric KinaseAssays

Compounds were tested further for activity against other proteinkinases. Protein kinase inhibition IC₅₀ data were determined usingstandardized radiometric kinase assays for each individual kinase, whichentail filter binding of ³³P labeled substrate proteins by the kinase ofinterest. Each IC₅₀ value was determined over a range of 10 drugconcentrations. Reaction conditions are available from the World WideWeb URL upstate.com/discovery/services/ic50_profiler.q.

KINASE

PIM1 46:35 108 40 PIM2 599 >1,000 66 PIM3 204 >1,000 58 CK2 alpha 1  396 102 CK2 alpha 2  1  1 78  39 DYRK2  91 354 138 FLT3(D835Y)  1 FLT3 35 721  9 HIPK2  86 LCK 240 MELK 184 CDK1/  56 226 cyclinB RAF1 238FLT4 316 815 GSK3B 512 HIPK3  45  56 RPS6KA1 390 DAPK3  17

The following kinase inhibition data were determined using standardizedradiometric kinase assays for each individual kinase, which entailfilter binding of ³³P labeled substrate proteins by the kinase ofinterest. Each percentage of activity was determined at 0.5 μMconcentration of the drug. Reaction conditions are available at theWorld Wide Web URL upstate.com/discovery/servi ce s/ic50_profiler.q.

KINASE

ABL1 9 7 20 14 ALK −12 −4 −20 −2 ARK5 −8 −18 14 5 ASK −16 −18 −1 2 AURKA6 −3 3 11 Blk(m) 0 14 33 BMX −18 −4 8 BRK −10 10 18 CAMK1 −4 −2 1 4CDK1/cyclinB 48 86 84 63 CDK2/cyclin A 37 60 53 53 CDK6/cyclinD3 −8 3 126 CDK7/cyclinH/ 23 42 36 57 MAT1 CDK9/cyclinT1 0 24 27 45 CHK1 1 12 13−1 CK1 gamma 1 −5 8 10 7 CK1 gamma 2 −7 19 35 −5 CK1 gamma 3 0 24 32 −5CK2 alpha 1 102 112 97 84 CK2 alpha 2 107 103 100 96 cKit(h) −14 2 15−10 cKit(D816H) −1 40 87 63 cKit(V560G) −11 19 69 75 RAF1 31 72 62 62CSK −46 −32 −9 14 DDR2 −9 −4 12 5 DRAK1 38 73 65 −5 DYRK2 50 95 55 82eEF-2K(h) −6 3 −8 −3 EGFR −23 −2 24 15 EGFR(L858R) 11 56 24 63EGFR(L861Q) 21 56 59 70 EGFR(T790M) 8 15 16 43 EGFR(T790M, −21 5 26 48L858R) EPHA5 −25 −35 12 8 EPHA7 1 2 5 0 EPHB4 −44 −31 −8 −16 ERBB4 −26−1 17 33 FAK 3 −13 −2 4

KINASE

FER −3 −17 6 8 FES −39 −33 1 −2 FGFR1 −3 16 23 17 FGFR2 −7 0 11 9 FLT1 528 75 19 FLT3(D835Y) 83 91 97 99 FLT3 58 82 90 100 FLT4 101 81 101 40CSF1R −74 −3 −12 52 FYN −14 18 18 32 GSK3B 44 55 28 26 HCK −11 25 26 28HIPK2 89 85 96 89 HIPK3 90 93 91 57 IGF1R 27 21 −9 −23 IKK alpha −1 −2 3−13 INSR −5 −6 −7 0 IRAK4 −19 −14 4 12 JAK2 1 2 38 −4 VEGFR2 33 61 55 15LCK 37 58 33 79 LOK 16 78 72 56 LYN 8; −9 21; 13 20 16 ERK2 5 6 21 15MAPKAPK2 −7 3 −12 −2 MEK1 −36 7 4 8 MELK 51 71 77 73 MER 54 82 86 62 MET−22 −21 −16 −6 MAP2K7 beta −33 −32 7 12 MLK1 20 43 21 49 Mnk2 37 79 −232 MSK2 44 34 41 9 MST1 4 20 −3 15 NEK2 23 66 73 13 p70S6K 20 32 36 8PAK2 −12 −12 1 4 PDGFRA −9 −6 −2 5 PDGFRA(D842V) −9 17 78 64

KINASE

PDGFRB −10 −2 −2 3 PDK1 −10 −9 8 7 PIM1 73 94 75 18 PKA −6; 10 2; 22 −9−1 AKT1 −4 1 7 7 PRKCA 1 0 9 1 PRKCT −11 −3 10 1 PRKd_nM2 −7 −4 0 25PRKG1 −5 1 −4 4 PLK3 −6 3 −1 0 MAPKAPK5 7 1 22 22 ROCK−I 3 4 12 11 RON−6 −6 9 −3 ROS −10 −8 5 −3 TYRO3 −9 14 22 2 RPS6KA1 22 60 54 55 PLK2 −1712 30 9 Src(1-530) −1 16 11 SRPK1 34 31 63 7 TAK1 −1 4 6 12 TIE2 1 2 −1245 TRKA 10 76 56 62 YES −9 18 34 30 ZAP70 −18 −8 2 −2 DAPK3 88 93 87 34ABL1(T315I) −7 0 ALK4 −15 −27 ABL2 2 7 AXL 16 59 BRSK1 −1 −3 BRSK2 7 15BTK −5 −8 CAMK2B 11 14 CAMK2G 36 40 CAMK2D 41; 6 40; 22 CAMK4 2 3CDK2/cyclinE −13 0 CDK3/cyclinE −19 −3 CDK5/p25 4 34

KINASE

CDK5/p35 −7 26 CHK2 5 20 CHK2(I157T)(h) 3 15 CHK2(R145W)(h) 0 6 CK1delta −4 5 cKit(D816V) 2 17 cKit(V654A) 8 9 CLK3 83 103 SRC −2 1 22DAPK1 58 77 DAPK2 91 94 DCAMKL2 −3 2 DMPK 4 3 EPHA1 −6 7 EPHA2 2 16EPHA3 −6 2 EPHA4 −3 10 EPHA8 −15 −10 EPHB1 1 21 EPHB2 13 29 EPHB3 −36−24 FGFR1(V561M) 12 16 FGFR2(N549H) 4 18 FGFR3 −2 3 FGFR4 14 14 FGR 3938 GCK 1 26 GRK5 11 74 GRK6 −2 44 GSK3A 51 56 GSG2 4 45 HIPK1 90 89 IKKbeta 5 13 IRAK1 4 32 INSRR −21 −18 ITK; Itk(h) 0 4 JAK3 8 37 JNK1A1 −9−7 JNK2A2 −7 −12

KINASE

JNK3 −3 4 LIMK1 3 4 LKB1 −2 19 MAPK2 −4 −1 MAPK2(m) −6 −7 MAPKAPK3 −8 5MARK1 0 6 MINK −13 −7 MKK4(m) −2 −17 MEK6 6 11 MLCK −3 −4 MRCKA −10 −19MRCKB 0 −4 MSK1 19 16 MSSK1 −3 11 MST2 −6 −8 MST3 0 17 MUSK −6 3 NEK11−2 0 NEK3 −12 −9 NEK6 −3 −1 NEK7 −14 −20 NLK 3 24 PAK3 21 16 PAK4 −12−12 PAK5 −8 −10 PAK6 −14 −1 PAR-1B alpha 7 8 PASK 87 85 PDGFRB(V561D) −413 PIM2 25 39 6 PIM3 13 55 −2 AKT2 −10 −11 AKT3 0 −5 PRKCB1 0 2 PRKCB2 18 PRKCG 0 6 PRKCD 7 0 PRKCE 0 −9

KINASE

PRKCZ −9 −12 PRKCN 2 −3 PRKCI −5 −4 PRKCM −4 −5 PRKG2 2 −10 PRK2 3 8PRKX 3 −2 FRK 3 5 Pyk2 −3 −1 RET 14 38 35 RIPK2 2 26 ROCK-II −4 1RPS6KA3 34 67 53 RPS6KA2 37 65 59 RPS6KA6 22 80 65 p38-alpha 1 33 p38-−8 −3 alpha(T106M) p38-beta 5 −1 p38-gamma 12 21 p38-delta −1 7 SGK −1 9SGK2 2 4 SGK3 2 −4 SLK −15 −10 SRPK2 36 34 STK33 0 67 SYK −9 10 TAO2 314 TAO3 22 57 TBK1 70 97 TLK2 8 35 TRKB 10 24 TSSK1 −18 −12 TSSK2 −9 −11VRK2 −3 −3 WNK2 11 4 WNK3 −17 15 mTOR 15 PLK1 −1

Example 12 Synthetic Processes Process 1

Methyl 5-(3-chlorophenylamino)benzo[c][2,6]naphthyridine-8-carboxylate(47 mg, 0129 mmol) was suspended in a mixture of methanol (1 ml) andhydrazine hydrate (1 ml). 3 drops of DMF were added and the mixturestirred at 60-70° C. for 2 hours. The volatiles were removed in vacuo.The resulting material was suspended in AcOEt/Hexanes, filtered anddried to afford5-(3-chlorophenylamino)benzo[c][2,6]naphthyridine-8-carbohydrazide assolid (47 mg, 100% yield). LCMS (ES): 95% pure, m/z 364 [M+1]⁺.

Process 2

5-(3-chlorophenylamino)benzo[c][2,6]naphthyridine-8-carbohydrazide (1.0eq, 21 mg, 0.057 mmol) was suspended in triethyl orthoformate (0.5 ml)and the mixture reacted in a microwave reactor at 120° C. for 80minutes. The precipitate that formed upon cooling was filtered and driedto affordN-(3-chlorophenyl)-8-(1,3,4-oxadiazol-2-yl)benzo[c][2,6]naphthyridin-5-amineas a solid (12 mg, 56% yield). LCMS (ES): 95% pure, m/z 374 [M+1]⁺.

Process 3

5-(3-chlorophenylamino)benzo[c][2,6]naphthyridine-8-carboxamide (1.0 eq,36 mg) was stirred in N,N-dimethylformamide dimethyl acetal (2 ml) at80° C. for 4 hours. The volatiles were removed in vacuo. Acetic acid wasadded (0.5 ml) and hydrazine hydrate (0.1 ml). The mixture was stirredat 80° C. for 1 hour. Water was added and the solid filtered and tried.After trituration in a mixture of CH₂Cl₂ and hexanes,N-(3-chlorophenyl)-8-(4H-1,2,4-triazol-3-yl)benzo[c][2,6]naphthyridin-5-aminewas isolated as a solid (22 mg, 67% yield). LCMS (ES): 95% pure, m/z 373[M+1]⁺.

Process 4

Methyl 3-bromoisonicotinate (1.0 eq, 1.76 g, 7.65 mmol),2-aminophenylboronic acid hydrochloride (1.0 eq, 1.33 g, 7.67 mmol) andcesium carbonate (2.0 eq, 4.99 g, 15.31 mmol) were suspended in dioxane(15 ml). The mixture was degassed by bubbling nitrogen for 10 minutes.PdCl₂(dppf) (0.05 eq, 280 mg, 0.383 mmol) was added and the mixture wasstirred at reflux for 2 hours. The resulting solid was filtered, washedwith methanol, water and methanol and dried.Benzo[c][2,6]naphthyridin-5(6H)-one was isolated as an off-white solid(823 mg, 55% yield). LCMS (ES): 95% pure, m/z 197 [M+1]⁺.

Process 5

Benzo[c][2,6]naphthyridin-5(6H)-one (1.0 eq, 813 mg, 4.15 mmol) wasstirred in phosphorus oxychloride (5.0 eq, 2 ml, 21.84 mmol) andacetonitrile (10 ml). The mixture was stirred at reflux for 5 hours. Themixture was poured on ice, and the resulting solid filtered and dried.5-chlorobenzo[c][2,6]naphthyridine was isolated as a grey solid (459 mg,52% yield). LCMS (ES): 95% pure, m/z 215 [M+1]⁺.

Process 6

2-methyl-5-nitrobenzoic acid (24 g) was dissolved in methanol (240 ml)and concentrated sulfuric acid (8 ml). The mixture was stirred at refluxovernight. Upon cooling the ester crystallized out. The material wasisolated by filtration to afford methyl 2-methyl-5-nitrobenzoate as awhite solid (19.0 g, 74% yield). A second crop of material (4.92 g, 19%yield) was isolated upon concentration and addition of water to themother liquor.

Methyl 2-methyl-5-nitrobenzoate (5.06 g) was suspended in methanol (100ml). The mixture was degassed by bubbling nitrogen for 15 minutes. Pd/C10% wet Degussa type E101 NE/WW (260 mg) was added and the mixturestirred under hydrogen atmosphere (balloon) overnight. The suspensionwas filtered and the solvents evaporated to afford methyl5-amino-2-methylbenzoate as an orange oil (4.18 g, 97% yield).

Methyl 5-amino-2-methylbenzoate (1.0 eq, 3.75 g) was dissolved in aceticacid (70 ml). N-Iodosuccinimide (1.0 eq, 5.27 g) was added portionwiseover 60 minutes. The mixture was stirred at room temperature for 30minutes. Acetic acid was evaporated. The residue was diluted with ethylacetate (80 ml) and neutralized with saturated sodium carbonate (80 ml).The organic layer was washed with 1M sodium thiosulfate (2×40 ml), thenwater (2×40 ml) and brine (2×40 ml). The material was purified by flashchromatography on silica gel (gradient 10% to 30% ethyl acetate inhexanes) to provide methyl 5-amino-4-iodo-2-methylbenzoate as ayellow-orange solid (3.19 g, 49% yield). GCMS >95% pure, adz 291. ¹H NMR(400 MHz, DMSO, d⁻⁶) δ 2.30 (s, 3H), 3.78 (s, 3H), 5.27 (br s, 2H), 7.24(s, 1H), 7.54 (s, 1H).

Process 7

2-amino-3-bromobenzoic acid (1.00 g) was mixed with methanol (10 ml) andconcentrated sulfuric acid (1 ml). The mixture was stirred at reflux for31 hours. The solvent were evaporated, and saturated aqueous sodiumbicarbonate was carefully added. The solid was extracted with CH₂Cl₂(3×). The combined extracts were dried over Na₂SO₄ and the solventsremoved in vacuo to afford methyl 2-amino-3-bromobenzoate as asemi-crystalline solid (976 mg, 91% yield). LCMS (ES): >85% pure, m/z230 [M+1]⁺.

Process 8

Methyl 2-amino-3-bromobenzoate (1.0 eq, 652 mg, 2.61 mmol) and4-(diisopropylcarbamoyl)pyridin-3-ylboronic acid (prepared according tothe procedure described in PCT patent application WO2005/105814), 1.0eq, 600 mg, 2.61 mmol) were combined with cesium carbonate (2.0 eq,1.699 g, 5.21 mmol) in dioxane containing 5% of water (6 ml). Themixture was degassed by bubbling nitrogen for 10 minutes. PdCl₂(dppf)(0.05 eq, 95 mg) was added and the reaction stirred at reflux for 2hours. Dioxane was evaporated, water was added and the materialextracted with CH₂Cl₂ (3×). The combined extracts were dried over Na₂SO₄and the solvents removed in vacuo. The material was purified by flashchromatography on silica gel (eluant 0.5% MeOH in CH₂Cl₂) to affordmethyl 2-amino-3-(4-(diisopropylcarbamoyl)pyridin-3-yl)benzoate as agreenish foam (244 mg, 31% yield). LCMS (ES): >95% pure, m/z 356 [M+1]⁺.

Process 9

Methyl 2-amino-3-(4-(diisopropylcarbamoyl)pyridin-3-yl)benzoate (1.0 eq,244 mg, 0.686 mmol) was dissolved under nitrogen atmosphere in anhydrousTHF (1.5 ml). A NaHMDS solution (1.0 M in THF, 2.0 eq, 1.4 ml, 1.4 mmol)was added dropwise through syringe. The resulting suspension was stirredat room temperature for 1 hour. The reaction was quenched by addition ofa saturated aqueous solution of ammonium chloride. The solid that formedwas filtered and dried. After trituration in methanol and filtration,methyl 5-oxo-5,6-dihydrobenzo[c][2,6]naphthyridine-7-carboxylate wasisolated as a grey fluffy solid (93 mg, 53% yield). LCMS (ES): >95%pure, m/z 255 [M+1]⁺.

The molecules in the following table were prepared using a similar twostep procedure from 4-(diisopropylcarbamoyl)pyridin-3-ylboronic acid andsuitable 2-iodo or 2-bromo amines:

TABLE 27 LCMS(ES) m/z, Structure MW [M + 1]+

316.35 317

197.19 198

197.19 198

214.20 215

280.20 281

221.21 222

268.27 269

254 255

Methyl 5-oxo-5,6-dihydrobenzo[c][2,6]naphthyridine-7-carboxylate (1.0eq, 85 mg, 0.334 mmol) was stirred in phosphorus oxychloride (2 ml) at120° C. for 2 hours. The solvent was removed in vacuo. Ice and waterwere added. The resulting solid was filtered and dried to afford methyl5-chlorobenzo[c][2,6]naphthyridine-7-carboxylate as a solid (84 mg, 92%yield). LCMS (ES): >95% pure, m/z 273 [M+1]⁺.

The molecules in the following table were prepared using a similarprocedure:

TABLE 28 LCMS(ES) m/z, Structure MW [M + 1]+

215.64 216

298.65 299

286.71 287

272.69 273

Process 11

Methyl 5-chlorobenzo[c][2,6]naphthyridine-7-carboxylate (1.0 eq, 48 mg,0.176 mmol) and 3-chloroaniline (3.0 eq, 60 ul, 0.56 mmol) were stirredunder microwave heating at 120° C. in NMP (0.3 ml) for 10 minutes. Waterwas added and the solid isolated by filtration. Trituration in methanoland filtration afforded methyl5-(3-chlorophenylamino)benzo[c][2,6]naphthyridine-7-carboxylate as asolid (29 mg, 45% yield). LCMS (ES): >85% pure, m/z 364 [M+1]⁺.

5-(3-chlorophenylamino)benzo[c][2,6]naphthyridine-7-carboxylate (29 mg)was stirred in ethanol (2 ml) and 6N aqueous NaOH (1 ml) at 60° C. for30 minutes. Water and HCl were added for reach pH=1. The resultingprecipitate was filtered, washed with water and dried to afford. LCMS(ES): >95% pure, m/z 350 [M+1]⁺.

The molecules in the following table were prepared using a similarprocedure.

TABLE 29 LCMS(ES) Structure MW m/z [M + 1]+

329.35 330

343.38 344

395.81 396

381.79 382

353.37 354

347.34 348

361.37 362

377.82 378

363.80 364 LCMS (ES) Structure MW [M + 1]+

349.77 350

329.35 330

315.33 316

Process 12

5-(3-chlorophenylamino)benzo[c][2,6]naphthyridine-7-carboxylic acid (20mg) was reacted in NMP (0.4 ml) with HOBt.H₂O (40 mg), ammonium chloride(40 mg), DIEA (100 ul) and EDCI (50 mg) at 70° C. for 1 hour. Water wasadded and the precipitate filtered and dried. After trituration inmethanol and filtration,5-(3-chlorophenylamino)benzo[c][2,6]naphthyridine-7-carboxamide wasisolated as a solid (8 mg). LCMS (ES): >95% pure, m/z 349 [M+1]⁺.

Process 13

5-chloropyrido[4,3-c][1,7]naphthyridine (10 mg) was mixed in NMP (0.3ml) with 3-chloroaniline (60 ul) and the mixture was heated at 120° C.for 10 min. Water was added and the resulting solid was filtered anddried. N-(3-chlorophenyl)pyrido[4,3-c][1,7]naphthyridin-5-amine wasisolated as a solid (5 mg). LCMS (ES)>95% pure, m/z 307 [M+1]⁺.

The molecules in the following table were prepared using a similarprocedure.

TABLE 30 LCMS (ES) m/z Structure MW [M + 1]+

272.30 273

302.33 303

306.75 307

336.78 337

286.33 287

300.36 301

316.36 317

315.33 316

329.36 330

307.39 308

292.26 293

Process 14

A solution of methyl 4-chloronicotinate (1.68 g, 6.05 mmol),2-amino-4-(methoxycarbonyl)phenylboronic acid hydrochloride (3.17 g,13.70 mmol), Cs₂CO₃ (8.90 g, 27.32 mmol), and PdCl₂(dppf) (335 mg, 0.46mmol) in dioxane (5% H₂O, 60 mL) was heated at reflux for 40 min. Thereaction was cooled to rt, the precipitate was collected by filtration,and washed (dioxane, H₂O, then with MeOH) to yield the desired lactam(2.07 g, 90%). LCMS (ES): >95% pure, m/z 255 [M+1]⁺.

Process 15

A solution methyl5-oxo-5,6-dihydrobenzo[c][2,7]naphthyridine-8-carboxylate (650 mg, 2.56mmol) in POCl₃ (4.0 mL) was heated at 120 C for 2.5 h. The reaction wasconcentrated under reduced pressure and diluted with ACN (20 mL) and H₂O(40 mL). The solution was neutralized with NaOH (3N) and the resultingprecipitate was collected by filtration to give the desired chloride(600 mg, 86%). LCMS (ES): >95% pure, m/z 273 [M+1]⁺.

A solution methyl 5-chlorobenzo[c][2,7]naphthyridine-8-carboxylate (60mg, 0.22 mmol) and 3-chloroaniline (50 uL) in NMP (1.0 mL) was heated at80 C for 1 h. Aqueous NaOH (3N, 0.3 mL) was added and continued heatingfor additional 30 min. The reaction was cooled to rt and added HCl (1N)until precipitate formed. The solid was collected by filtration andwashed with ACN to yield desired product (50 mg, 77%). LCMS (ES): >95%pure, m/z 350 [M+1]⁺.

The molecules in the following table were prepared using a similarprocedure.

TABLE 31 LCMS(ES) Structure MW m/z, [M + 1]+

349.77 350

367.76 368

315.33 316

349.77 350

367.76 368

339.35 340

315.33 316

343.38 344

333.32 334

343.38 344

367.76 368

315.33 316

329.35 330

333.32 334

339.35 340

350.76 351

368.75 369

316.31 317

344.37 345

334.30 335

330.34 331

340.33 341

Process 16

Methyl-3-bromothiophene carboxylate (1.0 eq, 2.42 g, 10.95 mmol),2-amino-4-cyanophenylboronic acid hydrochloride (1.05 eq, 2.28 g, 11.49mmol) and cesium carbonate (2.0 eq, 7.13 g, 21.9 mmol) were suspended indioxane (25 ml) containing 5% water. The mixture was degassed bybubbling nitrogen for 10 minutes. PdCl₂(dppf) (0.05 eq, 400 mg, 0.55mmol) was added and the mixture was stirred at reflux for 1.5 hours. Themixture was cooled down, the solid filtered, washed with dioxane, waterand methanol. After drying in vacuo,4-oxo-4,5-dihydrothieno[2,3-c]quinoline-7-carbonitrile was isolated as asolid (1.81 g, 73% yield). LCMS (ES) m/z 227 [M+1]⁺.

The molecules in the following table were prepared using a similarprocedure:

TABLE 32 LCMS (ES) m/z Structure MW [M + 1]+

226.3 227

259.0 260

Process 17

4-oxo-4,5-dihydrothieno[2,3-c]quinoline-7-carbonitrile (1.0 eq, 1.22 g,5.40 mmol) was stirred under reflux in acetonitrile (12 ml) andphosphorus oxychloride (5.0 eq, 2.5 ml, 26.8 mmol) for 6 hours. Thevolatiles were removed in vacuo, water and ice were added. The resultingsolid was filtered, washed with water and dried to afford4-chlorothieno[2,3-c]quinoline-7-carbonitrile as a light brown solid(1.18 g, 90% yield). LCMS (ES)>95% pure, m/z 245 [M+1]⁺.

The molecules in the following table were prepared using a similarprocedure:

TABLE 33 LCMS (ES) m/z Structure MW [M + 1]+

244 245

277 278

Process 18

4-chlorothieno[3,2-c]quinoline-7-carbonitrile (1.0 eq, 23 mg, 0.094mmol), aniline (0.1 ml) and NMP (0.1 ml) were mixed in a vial. Themixture was heated in a microwave oven at 120° C. for 10 um. Water wasadded and the resulting solid4-(phenylamino)thieno[3,2-c]quinoline-7-carbonitrile was filtered anddried. LCMS (ES): 95% pure, m/z 302 [M+1]⁺. This material was mixed in avial with DMF (0.5 ml), NH₄Cl (50 mg) and NaN₃ (50 mg). The mixture wasstirred at 120° C. for 3 hours. After addition of water and filtration,N-phenyl-7-(1H-tetrazol-5-yl) thieno[3,2-c]quinolin-4-amine was isolatedas a beige solid (13 mg, 41% yield). LCMS (ES): 95% pure, m/z 345[M+1]⁺, 317 [M+1−N₂]⁺. ¹H NMR (DMSO-d₆, 400 MHz) δ 7.07 (t, J=7.2, 1H),7.40 (t, J=7.6, 2H), 8.00 (dd, J=1.6, J=8.4, 1H), 8.04 (d, J=5.2, 1H),8.10 (dd, J=1.2, J=8.8, 2H), 8.19 (d, J=8.0, 1H), 8.25 (d, J=5.6, 1H),8.43 (d, J=1.6, 1H), 9.34 (s, 1H) ppm.

Process 19

4-chlorothieno[3,2-c]quinoline-7-carboxylate (10 mg, 0.036 mmol) wassuspended in NMP (0.1 ml) and 3-aminomethylpyridine (0.1 ml). Themixture was heated in a microwave oven at 120° C. for 10 nm. Thereaction mixture was dissolved in a mixture of NMP and MeOH and theester intermediate purified by preparative HPLC. After genevacevaporation of the solvents, the resulting solid was dissolved in a 1:1mixture of THF and MeOH (0.6 ml). 5N aqueous LiOH (0.2 ml) was added andthe mixture stirred at room temperature for 17 hrs. Water and aqueousHCl were added and the solution of4-(pyridin-3-ylmethylamino)thieno[3,2-c]quinoline-7-carboxylic acid waspurified by preparative HPLC. Removal of the solvents by genevacevaporation provided compound4-(pyridin-3-ylmethylamino)thieno[3,2-c]quinoline-7-carboxylic acid as awhite solid (10 mg, 62% yield). LCMS (ES): 95% pure, m/z 336 [M+1]⁺. ¹HNMR (CDCl₁, 400 MHz) δ 5.23 (s, 2H), 7.71-7.78 (m, 4H), 8.11 (d, J=5.6,1H), 8.47 (d, J=8.0, 1H), 8.49 (d, J=0.8, 1H), 8.62 (d, J=5.2, 1H), 8.97(s, 1H) ppm.

The molecules in the following table were prepared using a proceduresimilar to processes 18 and 19, using the appropriate startingmaterials.

TABLE 34 LCMS (ES) m/z, Structure MW [M + 1]+

335.81 336

319.36 320

319.36 320

335.81 336

353.80 354

369.36 370

396.83 397

362.38 363

378.84 379

396.83 397

378.84 379

362.38 363

380.37 381

412.39 413

336.45 337

296.39 297

336.45 337

339.42 340

379.48 380

379.48 380

356.35 357

388.36 389

338.36 339

354.81 355

370.37 371

402.39 403

352.38 353

362.38 363

380.37 381

378.84 379

396.83 397

412.39 413

396.83 397

378.84 379

362.38 363

336.45 337

408.52 409

296.39 297

336.45 337

379.48 380

351.43 352

339.42 340

379.48 380

Process 20

4-chlorothieno[3,2-c]quinoline-7-carbonitrile (1.0 eq, 23 mg, 0.094mmol), aniline (0.1 ml) and NMP (0.1 ml) were mixed in a vial. Themixture was heated in a microwave oven at 120° C. for 10 nm. Water wasadded and the resulting solid4-(phenylamino)thieno[3,2-c]quinoline-7-carbonitrile was filtered anddried. LCMS (ES): 95% pure, m/z 302 [M+1]⁺.

The molecules in the following table were prepared using a similarprocedure.

TABLE 35 LCMS (ES) m/z, Structure MW [M + 1]+

296.39 297

301.37 302

322.43 323

338.43 339

338.43 339

269.32 270

322.43 323

322.43 323

345.42 346

318.40 319

322.43 323

333.41 334

319.38 320

319.38 320

336.45 337

378.49 379

322.43 323

358.42 359

310.42 311

324.44 325

310.42 311

364.51 365

336.45 337

336.45 337

362.49 363

316.38 317

330.41 331

344.43 345

356.44 357

371.46 372

308.36 309

336.41 337

336.41 337

322.38 323

308.40 309

294.37 295

408.52 409

308.40 309

394.49 395

294.37 295

322.43 323

394.49 395

294.37 295

382.48 383

282.36 283

408.52 409

308.40 309

422.54 423

322.43 323

333.45 334

283.35 284

311.40 312

356.44 357

321.32 322

307.29 308

319.36 320

Process 21

To a solution of potassium t-butoxide (59 mg, 0.53 mmol) and EtOH washedRaney-Nickel in EtOH (50 mL) was added4-(3-fluorophenylamino)thieno-[3,2-c]quinoline-7-carbonitrile (560 mg,1.75 mmol) in EtOH (5 mL). The reaction mixture was charged with H₂ andstirred at rt for 3 h. Raney-Nickel was removed by filtration throughCelite and the solvent was removed under reduced pressure. Triturationin Et₂O gave the desired amine (300 mg, 53%) as a white solid. LCMS(ES): >95% pure, m/z 324 [M+1]⁺.

Process 22

To a solution of7-(aminomethyl)-N-(3-fluorophenyl)thieno[3,2-c]quinolin-4-amine (30 mg,0.09 mmol) in DCM (2 mL) was added phenyl isocyanate (10 uL, 0.09 mmol).The precipitate immediately appeared and it was collected by filtrationto give the desired urea as a white solid. LCMS (ES): >95% pure, m/z 443[M+1]⁺.

The molecules in the following table were prepared using a similarprocedure from corresponding amine and either isocyanate or chloride.

TABLE 36 LCMS (ES) m/z, [M + Structure MW 1]+

326.46 327

445.58 446

480.02 480

476.95 477

456.53 457

401.48 402

455.45 456

441.52 442

419.40 420

503.63 504

477.57 478

514.49 515

461.51 462

474.55 475

Process 23

4-(2-(dimethylamino)ethylamino)thieno[3,2-c]quinoline-7-carboxylic acid(1.0 eq, 100 mg) was mixed with ammonium chloride (2.0 eq, 34 mg), DIEA(114 ul), HOBt.H₂O (2.0 eq, 86 mg), EDCI (2.0 eq, 122 mg) in NMP (3 ml).The mixture was stirred at 70° C. until LCMS monitoring indicated acomplete reaction. Water was added, the pH was adjusted to 10 and thematerial was extracted with CH₂Cl₂. After evaporation of the solvents,the material was purified by preparative HPLC. Genevac evaporationafforded the TFA salt of4-(2-(dimethylamino)ethylamino)thieno[3,2-c]quinoline-7-carboxamide asyellow solid (92 mg, 69% yield). LCMS (ES)>95% pure, m/z 315 [M+1]⁺.

The molecules in the following table were prepared using a similarprocedure.

TABLE 37 LCMS (ES) m/z Structure MW [M + 1]+

432.54 433

418.51 419

417.53 418

417.53 418

412.55 413

410.53 411

366.48 367

356.44 357

420.53 421

420.53 421

328.43 329

354.47 355

342.46 343

314.41 315

396.55 397

372.48 373

Process 24

4-oxo-4,5-dihydrothieno[3,2-c]quinoline-7-carbonitrile (1.0 eq, 350 mg,1.55 mmol) was mixed with N-bromosuccinimide (1.1 eq, 303 mg, 1.70 mmol)in acetic acid (4 ml). The mixture was stirred at 100° C. for 4 hours.The mixture was cooled down to 80° C., more NBS (303 mg) was added andthe mixture stirred overnight. Water was added and the material filteredand dried. Trituration in methanol and filtration afforded2-bromo-4-oxo-4,5-dihydrothieno[3,2-c]quinoline-7-carbonitrile as a greysolid (396 mg, 84% yield). LCMS (ES)>80% pure, m/z 305[M]⁺, 307 [M+2]⁺.

This crude material was treated with phosphorus oxychloride (5.0 eq, 0.6ml, 6.33 mmol) in acetonitrile (4 ml) at reflux for 4 hours. More POCl₃(2 ml) was added and the mixture heated at 110° C. for 7 hours. Thevolatiles were removed, Ice was added and the solid filtered. Aftertrituration in ethyl acetate/hexanes and filtration,2-bromo-4-chlorothieno[3,2-c]quinoline-7-carbonitrile was isolated as asolid (324 mg, 78% yield). LCMS (ES)>80% pure, m/z 323[M]⁺, 325 [M+2]⁺.

2-bromo-4-chlorothieno[3,2-c]quinoline-7-carbonitrile (1.0 eq, 309 mg,0.955 mmol) and N,N-dimethylene diamine (3.0 eq, 312 ul, 2.85 mmol) weremixed in NMP (1 ml). The mixture was heated under microwave at 100° C.for 10 min. Water was added and the solid filtered. The material waspurified by trituration in hot ethyl acetate.2-bromo-4-(2-(dimethylamino)ethylamino)thieno[3,2-c]quinoline-7-carbonitrilewas isolate as a solid (252 mg, 70% yield). LCMS (ES)>95% pure, m/z375[M]⁺, 377 [M+2]⁺.

Process 25

2-bromo-4-(2-(dimethylamino)ethylamino)thieno[3,2-c]quinoline-7-carbonitrile(20 mg) was mixed with sodium azide (50 mg) and ammonium chloride (50mg) in DMF. The mixture was stirred at 120° C. for 3 hours. Water wasadded and the solid isolated by filtration. (6 mg). LCMS (ES)>95% pure,m/z 418[M]⁺, 420 [M+2]⁺.

Process 26

2-bromo-4-(2-(dimethylamino)ethylamino)thieno[3,2-c]quinoline-7-carbonitrile(1.0 eq, 55 mg, 0.146 mmol), benzene boronic acid (2.0 eq, 36 mg, 0.295mmol), cesium carbonate (2.0 eq, 95 mg, 0.292 mmol) and PdCl₂(dppf)(0.05 eq, 5 mg, 0.068 mmol) were mixed in dioxane (0.5 ml) containing 5%of water. The mixture was heated under microwave for 10 min at 120° C.After addition of water and filtration,4-(2-(dimethylamino)ethylamino)-2-phenylthieno[3,2-c]quinoline-7-carbonitrilewas isolated as a solid. LCMS (ES) m/z 373 [M+1]⁺.

This solid was dissolved in DMF (0.5 ml) and treated with sodium azide(100 mg) and ammonium chloride (100 mg) at 120° C. for 1.5 hours. Waterwas added and filtration of the solid providedN1,N1-dimethyl-N2-(2-phenyl-7-(1H-tetrazol-5-yethieno[3,2-c]quinolin-4-yl)ethane-1,2-diamine.(35 mg). LCMS (ES)>85% pure, m/z 416 [M+1]⁺.

Process 27

Phenol (2.0 eq, 85 mg) was dissolved in anhydrous DMF. 60% sodiumhydride (2.0 eq, 36 mg) was added and the reaction mixture stirred for afew minues. 4-chlorothieno[3,2-c]quinoline-7-carbonitrile (1.0 eq, 110mg) was added to the mixture and the whole reaction was stirred at 100°C. for two days. Water was added and the solid was filtered and dried.4-phenoxythieno[3,2-c]quinoline-7-carbonitrile was isolated as a solid(114 mg). LCMS (ES)>95% pure, m/z 303 [M+1]⁺.

Process 28

In an oven dried flask, under nitrogen atmosphere, was charged sodiumazide (1.4 eq, 84 mg). Et₂AlCl (1.4 eq, 1.08 ml of 1.8 M solution intoluene) was added through syringe. The mixture was stirred at roomtemperature for 4 hours. 4-phenoxythieno[3,2-c]quinoline-7-carbonitrile(1.0 eq, 20 mg) was charged in a vial. Et₂AlN₃ solution (0.15 ml) wasadded and the resulting mixture stirred at 80° C. for 5 days. Themixture was treated by a solution of NaOH and some sodium nitrite wasadded (pH=13-14). The pH was adjusted to 1.5 with HCl 6N. The materialwas extracted with ethyl acetate. The material was extracted from theorganic phase using a saturated aqueous solution of K₂CO₃. The pH wasadjusted to 2.5 with HCl 6N and the material was extracted with ethylacetate. The solvent were evaporated to afford4-phenoxy-7-(1H-tetrazol-5-yl)thieno[3,2-c]quinoline. LCMS (ES)>95%pure, m/z 303 [M+1]⁺.

The molecules in the following table were prepared using a proceduresimilar to process 27 and process 28.

TABLE 38 LCMS (ES)m/z, Structure MW [M + 1]+

302.35 303

308.28 309

316.38 317

297.37 298

359.40 360

340.40 341

351.31 352

338.33 339

353.40 354

320.34 321

344.39 345

336.79 337

345.38 346

363.37 364

379.82 380

Biological activities for various compounds are summarized in thefollowing table.

Lengthy table referenced here US20120208792A1-20120816-T00001 Pleaserefer to the end of the specification for access instructions.

Additional methods for preparing certain compounds of the invention,including compounds of Formula IA, IB and IC, are provided.

Process 29: Synthesis of Halogeno Aniline

Methyl 2-amino-5-fluorobenzoate (1.0 eq, 8.47 g, 0.051 mol) was reactedwith N-Iodo succinimide (1.03 eq, 11.6 g, 0.0515 mol) in acetic acid(100 ml) at room temperature for 20 minutes. The solvent was removed invacuo. A K₂CO₃ aqueous solution was added and the compound extractedwith ethylacetate. The organic layer was washed with 1M sodiumthiosulfate, water and then brine. After drying over Na₂SO₄, andevaporation of the solvent, methyl 2-amino-5-fluoro-3-iodobenzoatewasisolated as a purple solid (14.21 g, 96% yield). ¹H NMR (CDCl₃, 400 MHz)δ 3.8 (s, 3H), 6.3 (br s, 2H), 7.6 (m, 2H) ppm.

Process 30

The boronic ester was prepared in two steps using the proceduresdescribed by Alessi et al., J. Org. Chem., 2007, 72, 1588-1594.

Process 31

Step A

Methyl 2-amino-3-bromobenzoate (1.0 eq, 652 mg, 2.61 mmol) and4-(diisopropylcarbamoyl)pyridin-3-ylboronic acid (prepared according tothe procedure described in PCT patent application WO2005/105814, 1.0 eq,600 mg, 2.61 mmol) were combined with cesium carbonate (2.0 eq, 1.699 g,5.21 mmol) in dioxane containing 5% of water (6 ml). The mixture wasdegassed by bubbling nitrogen for 10 minutes. PdCl₂(dppf) (0.05 eq, 95mg) was added and the reaction stirred at reflux for 2 hours. Dioxanewas evaporated, water was added and the material extracted with CH₇Cl₂(3×). The combined extracts were dried over Na₂SO₄ and the solventsremoved in vacuo. The material was purified by flash chromatography onsilica gel (eluant 0.5% MeOH in CH₂Cl₂) to afford methyl2-amino-3-(4-(diisopropylcarbamoyl)pyridin-3-yl)benzoate as a greenishfoam (244 mg, 31% yield). LCMS (ES): >95% pure, m/z 356 [M+1]⁺.

Step B:

Methyl 2-amino-3-(4-(diisopropylcarbamoyl)pyridin-3-yl)benzoate (1.0 eq,244 mg, 0.686 mmol) was dissolved under nitrogen atmosphere in anhydrousTHF (1.5 ml). A NaHMDS solution (1.0 M in THF, 2.0 eq, 1.4 ml, 1.4 mmol)was added dropwise through syringe. The resulting suspension was stirredat room temperature for 1 hour. The reaction was quenched by addition ofa saturated aqueous solution of ammonium chloride. The solid that formedwas filtered and dried. After trituration in methanol and filtration,methyl 5-oxo-5,6-dihydrobenzo[c][2,6]naphthyridine-7-carboxylate wasisolated as a grey fluffy solid (93 mg, 53% yield). LCMS (ES): >95%pure, m/z 255 [M+11]⁺.

The following compounds were prepared using similar chemistries byreacting appropriate boronic esters and acids with appropriate2-halogenoanilines

TABLE 40 Structure M.W. LCMS (ES) m/z

254.24 255 [M + 1]⁺

272.23 273 [M + 1]⁺

272.23 273 [M + 1]⁺

Process 32

Methyl 5-oxo-5,6-dihydrobenzo[c][2,6]naphthyridine-7-carboxylate (1.0eq, 85 mg, 0.334 mmol) was stirred in phosphorus oxychloride (2 ml) at120° C. for 2 hours. The solvent was removed in vacuo. Ice and waterwere added. The resulting solid was filtered and dried to afford methyl5-chlorobenzo[c][2,6]naphthyridine-7-carboxylate as a solid (84 mg, 92%yield). LCMS (ES): >95% pure, m/z 273 [M+1]⁺.

The following compounds were prepared using similar chemistries on theappropriate lactams:

TABLE 41 Structure M.W. LCMS (ES) m/z

272.69 273[M + 1]⁺

290.68 291[M + 1]⁺

290.68 291[M + 1]⁺

Certain compounds of the present invention, including compounds ofFormula IA, IB and IC, can be prepared according to the followinggeneral processes, using appropriate materials.

The following molecules can be prepared using the chemistry of GeneralProcess 1:

The following molecules can be prepared using chemistry of GeneralProcess 2:

The following molecules can be prepared using chemistry of GeneralProcess 3:

Process 33

Step A:

Methyl 5-chlorobenzo[c][2,6]naphthyridine-7-carboxylate (1.0 eq, 48 mg,0.176 mmol) and 3-chloroaniline (3.0 eq, 60 ul, 0.56 mmol) were stirredunder microwave heating at 120° C. in NMP (0.3 ml) for 10 minutes. Waterwas added and the solid isolated by filtration. Trituration in methanoland filtration afforded methyl5-(3-chlorophenylamino)benzo[c][2,6]naphthyridine-7-carboxylate as asolid (29 mg, 45% yield). LCMS (ES): >85% pure, m/z 364 [M+1]⁺.

Step B:

5-(3-chlorophenylamino)benzo[c][2,6]naphthyridine-7-carboxylate (29 mg)was stirred in ethanol (2 ml) and 6N aqueous NaOH (1 ml) at 60° C. for30 minutes. Water and HCl were added for reach pH=1. The resultingprecipitate was filtered, washed with water and dried to afford5-(3-chlorophenylamino)benzo[c][2,6]naphthyridine-7-carboxylic acid as asolid. LCMS (ES): >95% pure, m/z 350 [M+1]⁺.

The following compounds were prepared using similar chemistries

TABLE 42 LCMS (ES) Structure MW m/z [M + 1]⁺

349.77 350

367.76 368

367.76 368

349.77 350

329.35 330

315.33 316

363.80 364

Process 34

5-(3-chlorophenylamino)benzo[c][2,6]naphthyridine-7-carboxylic acid (20mg) was reacted in NMP (0.4 ml) with HOBt.H₂O (40 mg), ammonium chloride(40 mg), DIEA (100 ul) and EDCI (50 mg) at 70° C. for 1 hour. Water wasadded and the precipitate filtered and dried. After trituration inmethanol and filtration,5-(3-chlorophenylamino)benzo[c][2,6]naphthyridine-7-carboxamide wasisolated as a solid (8 mg). LCMS (ES): >95% pure, m/z 349 [M+1]⁺.

The following compounds were prepared using similar chemistries byreacting the appropriate carboxylic acid and the appropriate substitutedor unsubstituted amines.

TABLE 43 LCMS (ES) m/z [M + Structure MW 1]⁺

332.33 333

348.79 349

332.33 333

348.79 349

388.42 389

366.78 367

419.91 420

406.86 407

362.81 363

376.84 377

438.91 439

418.88 419

388.85 389

424.88 425

378.81 379

Process 35

Methyl 5-(3-chlorophenylamino)benzo[c][2,6]naphthyridine-7-carboxylate(19 mg, 0.052 mmol) was suspended in anhydrous THF (0.5 ml). A 1.0 M THFsolution of LiAlH₄ (0.2 ml, 0.2 mmol) was added and the mixture wasstirred at room temperature for 3 hours. Another amount of LiAlH₄solution (0.3 ml, 0.3 mmol) was added and the mixture stirred at 60° C.for 45 min. Water was added and the mixture was stirred at roomtemperature overnight. Methanol was added and the mixture was filteredthrough celite. The solvent were evaporated. The material was purifiedby preparative TLC on silica gel (5% MeOH in CH₂Cl₂) and preparativeHPLC. Genevac evaporation afforded 4 mg of the TFA salt of(5-(3-chlorophenylamino)benzo[c][2,6]naphthyridin-7-yl)methanol as asolid. LCMS (ES): >90% pure, m/z 336 [M+1]⁺.

Process 36

Methyl 5-(3-chlorophenylamino)benzo[c][2,6]naphthyridine-7-carboxylate(47 mg) was mixed with Methanol (1 ml) and Hydrazine hydrate (1 ml). 2-3drops of DMF were added and the mixture was stirred at 60° C. for 2hours. The volatiles were removed and another amount of reagent Methanol(1 ml) and Hydrazine (1 ml) were added, and the mixture was stirred at60° C. for an extra 2 hours. The volatiles were removed in vacuo and thematerial was crashed out using AcOEt/hexanes. Filtration and dryingafforded5-(3-chlorophenylamino)benzo[c][2,6]naphthyridine-7-carbohydrazide as asolid (29 mg, 62% yield). LCMS (ES): >85% pure, m/z 364 [M+1]⁺.

Process 37

5-(3-chlorophenylamino)benzo[c][2,6]naphthyridine-7-carbohydrazide (24mg) was stirred in triethylorthoformate (1 ml) at 120° C. for 4 hours.The solid was filtered and triturated in CH₂Cl₂/MeOH. Impurities (mainlystarting material) were removed by filtration and the filtratecontaining the expecting compound was concentrated. The material waspurified by preparative TLC on silica gel (5% methanol in CH₂Cl₂) toaffordN-(3-chlorophenyl)-7-(1,3,4-oxadiazol-2-yl)benzo[c][2,6]naphthyridin-5-amineas a solid (8 mg). LCMS (ES): >95% pure, m/z 374 [M+1]⁺.

Process 38

5-(3-chlorophenylamino)benzo[c][2,6]naphthyridine-7-carboxamide (12 mg,0.034 mmol) was suspended in dichloroethane (0.2 ml). Sodium chloride(70 mg) was added followed by Phosphorus oxychloride (20 ul). Themixture was stirred at 80° C. for 1.5 hours. An extra amount ofPhosphorus oxychloride (50 ul) was added and the mixture was heated at80° C. for 8 hours. The volatiles were removed in vacuo. Water was addedand the solid was filtered. The material was purified by preparative TLCon silica gel (5% MeOH in CH₂Cl₂) to provide 6 mg of5-(3-chlorophenylamino)benzo[c][2,6]naphthyridine-7-carbonitrile. LCMS(ES): >95% pure, adz 331 [M+1]⁺.

Process 39

5-(2-chlorophenylamino)benzo[c][2,6]naphthyridine-7-carbonitrile (50 mg,0.151 mmol) was stirred in a vial at 120° C. for 2 hours in the presenceof DMF (0.5 ml), sodium azide (88 mg, 1.35 mmol) and ammonium chloride(72 mg, 1.35 mmol). Water was added, the pH was lowered and theresulting solid was filtered. The material was dissolved in NMP andpurified by preparative HPLC. Genevac evaporation afforded the TFA saltofN-(2-chlorophenyl)-7-(1H-tetrazol-5-yl)benzo[c][2,6]naphthyridin-5-amine(8 mg). LCMS (ES): >95% pure, m/z 374 [M+1]⁺, 346 [M+1−N2]⁺.

Process 40

5-(3-chlorophenylamino)benzo[c][2,6]naphthyridine-7-carboxamide (18.5mg) was stirred at 80° C. overnight in DMF-DMA (0.7 ml). The solvent wasevaporated. Hydrazine hydrate (1 ml) and acetic acetic (1 ml) were addedand the mixture stirred at 80° C. for one hour. Water was added and theresulting solid was filtered. The material was suspended in Methanol,filtered and dried to affordN-(3-chlorophenyl)-7-(4H-1,2,4-triazol-3-yl)benzo[c][2,6]naphthyridin-5-amineas a yellow solid (4.8 mg). LCMS (ES): >90% pure, m/z 373 [M+1]⁺.

Process 41

Methyl 5-oxo-5,6-dihydrobenzo[c][2,6]naphthyridine-7-carboxylate (2.17g, 8.53 mmol) was mixed with 6N aqueous sodium hydroxide (10 ml) andEthanol (40 ml). The mixture was stirred at reflux for 5 hours. Aftercooling down, water was added and the mixture was acidified by 6N HCl.After filtration and drying5-oxo-5,6-dihydrobenzo[c][2,6]naphthyridine-7-carboxylic acid wasisolated as a grey solid (1.91 g, 93% yield). LCMS (ES): mh 241 [M+1]⁺.

Process 42

5-oxo-5,6-dihydrobenzo[c][2,6]naphthyridine-7-carboxylic acid (1.0 eq,1.91 g, 7.96 mmol) was mixed with HOBt.H₂O (2.0 eq, 2.15 g, 15.91 mmol)and NH₄Cl (8.0 eq, 3.41 g, 63.6 mmol) in NMP (30 ml). DIEA (4.0 eq, 5.5ml, 31.57 mmol) and EDCI (2.0 eq, 3.05 g, 15.91 mmol) was added and themixture was stirred in a closed vessel at 80° C. for 2.5 hours. Waterand brine were added. The solid was filtered, washed with water, washedwith methanol and dried in a vacuum oven.5-oxo-5,6-dihydrobenzo[c][2,6]naphthyridine-7-carboxamide was isolatedas an off-white solid (1.81 g, 96% yield). LCMS (ES): m/z 240 [M+1]⁺.

Process 43

5-oxo-5,6-dihydrobenzo[c][2,6]naphthyridine-7-carboxamide (1.81 g, 7.59mmol) was stirred in DMF-DMA (20 ml) at 80° C. for 1.5 hours. Thevolatiles were removed in vacuo. This operation was repeated severaltimes until the amount of starting material detected by LCMS remainedconstant. After evaporation of the volatiles, the crude intermediate wassuspended in acetic acid (40 ml). Hydrazine hydrate (4 ml) was addeddropwise and the reaction mixture was stirred without externaltemperature control for about 10 minutes. The reaction was then stirredat 80° C. for 45 minutes upon which the mixture turned into a thickmass. Water was added and the solid filtered. After washing with waterand drying, 7-(4H-1,2,4-triazol-3-yl)benzo[c][2,6]naphthyridin-5(6H)-onewas isolated as a pale grey solid (1.84 g, 92% yield). LCMS (ES): m/z264 [M+1]⁺.

Process 44

Step A:

Under nitrogen atmosphere,7-(4H-1,2,4-triazol-3-yl)benzo[c][2,6]naphthyridin-5(6H)-one (1.8 g,6.84 mmol) was mixed with Phosphorus oxychloride (3.2 ml) inacetonitrile (20 ml). The mixture was stirred overnight at 100° C. Thevolatiles were removed in vacuo. The resulting solid was suspended inCH₂Cl₂ and a little bit of MeOH. After filtration and drying, crude5-chloro-7-(4H-1,2,4-triazol-3-yl)benzo[c][2,6]naphthyridine (2.04 g)was isolated as a greenish solid. LCMS (ES) m/z 282 [M+1]⁺.

Step B:

The product of step A (57 mg) was mixed with aniline (100 ul) in NMP(0.5 ml) and the mixture heated in a microwave oven at 120° C. for 10minutes. An additional NMP (1.5 ml) was added and the solution filtered.Purification by preparative HPLC and Genevac evaporation provided asolid that was further purified by trituration in AcOEt/hexanes. The TFAsalt ofN-phenyl-7-(4H-1,2,4-triazol-3-yl)benzo[c][2,6]naphthyridin-5-amine wasisolated as a solid (34 mg). LCMS (ES): >95% pure, m/z 339 [M+1]⁺.

The molecules depicted in the following table were prepared usingchemistries similar to the one described in Processes 40 and 44:

TABLE 44 LCMS (ES) m/z Structure MW [M + 1]⁺

372.81 373

390.80 391

370.38 371

390.80 391

368.39 369

374.35 375

352.39 353

374.35 375

356.36 357

338.37 339

382.42 383

356.36 357

370.38 371

302.33 303

362.39 363

372.81 373

372.81 373

368.39 369

368.39 369

382.42 383

370.38 371

374.35 375

390.80 391

372.81 373

363.37 364

390.80 391

386.84 387

366.42 367

316.36 317

374.35 375

417.44 418

386.38 387

422.36 423

356.36 357

386.84 387

320.35 321

306.32 307

406.36 407

412.44 413

412.44 413

422.36 423

353.38 354

353.38 354

373.45 374

353.38 354

390.80 391

345.40 346

375.43 376

333.39 334

373.45 374

390.80 391

388.81 389

456.81 457

444.87 445

330.39 331

332.36 333

316.36 317

386.38 387

316.36 317

367.41 368

443.48 444

470.96 471

430.43 431

469.51 470

425.49 426

459.93 460

501.97 502

450.90 451

467.52 468

436.51 437

The following molecules can be prepared using similar chemistry:

Further methods for preparing certain compounds of the invention,including compounds of Formula IA, IB and IC, are provided.

Process 45

2-chloro-4-fluoronitrobenzene (1.0 eq, 1 g, 5.7 mmol), N-methylpiperazine (1.2 eq, 1.18 g, 6.84 mmol), potassium carbonate (2.0 eq, 1.6g, 11.6 mmol) were stirred at 100° C. in DMF for 3 hours. The mixturewas cooled down and diluted with water. The material was extracted withethyl acetate. The organic layer was washed with brine, dried overNa₂SO₄ and the solvent evaporated in vacuo. After trituration indiethylether and filtration,1-(3-chloro-4-nitrophenyl)-4-methylpiperazine was isolated as a solid(0.9 g, 62%). LCMS (ES): m/z 256 [M+1]⁺. This material was suspended inMeOH (20 ml) with Raney Nickel (0.2 g) and stirred under hydrogenatmosphere overnight. The catalyst was filtered off through celite.Evaporation of the solvents provided2-chloro-4-(4-methylpiperazin-1-yl)aniline as a dark brown oil (0.68 g,86%). LCMS (ES): m/z 226 [M+1]⁺.

Process 46

2-chloro-4-(2-morpholinoethoxy) aniline was obtained in two steps from2-chloro-4-fluoronitrobenzene and 4-(2-hydroxyethyl) morphine using aprotocol described in patent application WO2008/42282.

Process 47

2-chloro-4-(2-(dimethylamino)ethoxy)aniline was prepared according tothe procedure described in General Process 2.

Process 48

2-fluoro-4-(2-(pyrrolidin-1-yl)ethoxy)aniline was prepared in two stepsusing a procedure described in patent application WO2007/7152.

Process 49

4-(2-(dimethylamino)ethoxy)-2-fluoroaniline was prepared in two stepsusing the procedure described in Process 46.

Process 50

2-fluoro-4-(2-methoxyethoxy)aniline was prepared in one step using aprocedure described in patent application US2006/155128.

Process 51

Step A:

4 amino-3-chlorobenzonitrile was charged in a vial. NaHMDS (1 M solutionin THF, 0.2 ml) was added and the solution stirred at 80° C. for 5 nm. Asuspension of5-chloro-7-(4H-1,2,4-triazol-3-yl)benzo[c][2,6]naphthyridine (30 mg) inNMP (0.5 ml) was added and the solution stirred at 80° C. for 30 min.The mixture was cooled down, a few drops of HCl and NMP (1 ml) wereadded and mixture was purified by preparative HPLC to provide4-(7-(4H-1,2,4-triazol-3-yl)benzo[c][2,6]naphthyridin-5-ylamino)-3-chlorobenzonitrile(25 mg).

Step B:

The material from step A was treated with LiAlH₄ (20 mg) in dry THF (1ml) and the solution stirred at 60° C. for several hours. The reactionwas then treated with Na₂SO₄.10.H₂O and filtered. The residue waspurified by preparative HPLC to affordN-(4-(aminomethyl)-2-chlorophenyl)-7-(4H-1,2,4-triazol-3-yl)benzo[c][2,6]naphthyridin-5-amineas a TFA salt (3 mg). LCMS (ES): >85% pure, m/z 402 [M+1]⁺, 385[M+1−NH₃]⁺.

Process 52

Reaction of 5-chloro-7-(4H-1,2,4-triazol-3-yl)benzo[c][2,6]naphthyridinewith organoboranes under conditions of the Suzuki reaction affordscompound m.

The following are examples of organoboranes that can be used in theSuzuki coupling reaction with5-chloro-7-(4H-1,2,4-triazol-3-yl)benzo[c][2,6]naphthyridine.

Process 57: Synthesis of5-phenyl-7-(4H-1,2,4-triazol-3-yl)benzo[c][2,6]naphthyridine

To 5-chloro-7-(4H-1,2,4-triazol-3-yl)benzo[c][2,6]naphthyridine (21.3mg), cesium carbonate (49 mg) and phenylboronic acid (19 mg) in dioxane(1 mL) was added PdCl₂(dppf) under nitrogen atmosphere. The mixturestirred at 120° C. at 300 W (microwave) for 10 min. Water was added andresidue obtained after extraction with dichloromethane was purified bypreparative HPLC. LCMS (ES) m/z [M+1]⁺324.

Biological data for representative compounds of the invention isprovided in Tables 45 and 46:

TABLE 45 Biological data. MV- PIM1: PIM2: phosphoBAD 4-11 K-562 MiaPaCaIC50 IC50 ser 112 IC50 IC50 IC50 IC50 MDAMB231 Structure (uM) (uM) (uM)(uM) (uM) (uM) IC50 (uM)

<0.1 <0.1 <0.1 <0.1 <5 <0.5 <0.5

<0.1 <0.1 <0.5 <0.5 <0.5 >10

<0.1 <0.1 <0.1 <0.1 <0.5 <5 >10

<0.1 <0.1 <1 <0.1 <5 <1 <1

<0.1 <0.1 <0.1 <0.1 <0.5 <0.5

<0.1 <0.1 <0.1 <0.1 <5 <5 <5

<0.1 <0.1 <0.1 <0.1 <1 <1 >10

<0.1 <0.1 <0.1 <0.1 <0.5 <1 <5

<0.1 <0.1 <5 <0.1 <0.5 <1

<0.1 <1 <0.1 <0.1 <5 <1 >10

<0.1 <0.5 <0.1 <0.1 <5 <1 >10

<0.1 <0.5 <0.5 <0.5 >10 <5 <5

<0.1 <0.1 <5 <0.1 >10 >10

<0.1 <0.1 <0.1 <0.1 <0.1 <1 <1

<0.1 <0.1 <0.1 <0.1 <5 <0.5 <0.5

<0.1 <0.1 <1 <0.5 <1 <1 <5

<0.1 <0.5 <5 <0.5 <0.5 >10 >10

<0.1 <1 <0.1 <0.5 <5 <1 <1

<0.1 <1 <0.1 <0.1 <5 v <5

<0.1 <0.5 <5 <0.5 <0.5 >10 >10

<0.1 <0.5 <5 <0.1 >10 <0.5 >10

<0.1 <0.5 <5 <0.1 >10 <5 <1

<0.1 <0.5 <1 <0.1 <5 <5 <5

<0.1 <0.5 >10 >10 >10 >10

<0.1 >1.1 <5 <0.1 >10 >10

<0.1 >1.1 <0.5 <0.1 <5 <0.5 <1

<0.1 <1 <5 <0.1 <5 >10

<0.1 <0.5 <1 <0.5 >10 >10 <10

<0.1 <0.5 <0.1 <0.1 >10 >10 v

<0.1 <0.5 <1 <0.5 <5 <1 >10

<0.1 <0.5 <1 <0.5 >10 >10 >10

<0.1 <0.5 <5 <0.5 <5 <10 >10

<0.1 <0.5 <0.1 <0.1 <5 <1 >30

<0.1 <0.1 <0.1 <0.5 >10 >10 >10

<0.1 <0.5 <0.5 v <1 v

<0.1 <5 <1 <0.5 <5 >10

<0.1 <0.5 <0.1 <0.1 <10 <5 <10

<0.1 <5 <0.1 <0.5 <1 <10

<0.5 <1 <0.5 <0.1 >10 <0.5 >10

<0.5 >1.1 <1 <0.5 <5 <5

<0.5 <0.1 <0.12 <1 <1 <1

<0.5 <1 <0.5 <0.1 >10 <1 <10

<0.5 >10 >10 >10 >10

<0.5 <0.1 <0.1 >10 >10 >10

<0.5 <1 <5 <0.5 <0.5 <5 >10

<0.5 <1

<0.5 <0.5

<0.5 >1.1

<0.5 >1.1

<0.5

<1 <1

<1 >1.1

<1 >1.1

<1

<5 >1.1

<5 >1.1

>1.1

>1.1 >1.1

>1.1 >1.1

>1.1 >1.1

>1.1 >1.1

>1.1

>1.1

>1.1

>1.1

>1.1

>1.1

>1.1

>1.1

>1.1

>1.1

>1.1

>1.1

>1.1

>1.1

>5

>1.1

>1.1

TABLE 46 Biological data. phospho FLT3 BAD MV- MDA Mia- PIM1 autophosser112 K-562 411 MB23 PaCa PC3 IC₅₀ IC₅₀ IC₅₀ IC₅₀ IC₅₀ 1 IC₅₀ IC₅₀ IC₅₀Structure (uM) (uM) (uM) (uM) (uM) (uM) (uM) (uM)

<0.1 >10 <0.1 9.638 >30

<0.1 <1 <0.1 2.179 11.983

<0.1 <1 <0.1 2.936 1.498 3.658

<0.1 <0.5 <0.1 <1 <1

<0.1 >10 <0.1 <0.5 <0.1 <1

<0.1 >10 <0.1 <0.1 <0.1 2.555 1.765

<0.1

<0.1 5.41 <0.1 6.939 <0.1 1.231

<0.1 6.011 <0.1 >10 <0.1 >10 >30

<0.1 2.655 <0.1 2.73 1.507

<0.1 >10 <0.5 <0.5 <0.1 2.036 0.268 <1

<0.5

<0.1 >10 <0.1 <0.5 <0.1 >10 7.625 <0.5

<0.1 >10 0.51 <1 <0.1 6.859 <0.5 <0.5

<0.5 >10 2.945 3.226 <1 >10 <0.1 >30

3.967

<0.1 >10 <0.5 1.926 <0.5 >10 >30

<0.1 >10 <0.1 >10 >10 >30

<0.1 >10 4.173 >10 <0.1 >10 >10 >30

<0.1 <0.5 >10 2.38 <1 <0.5

<0.1 >10 <0.1 >10 <0.5 >30

The following table is the %-activity data of compound A in differentkinase enzymes at 0.5 μM of ATP.

TABLE 47 Kinase % Activity DYRK2(h) −4 HIPK2(h) −1 Pim-1(h) 1 HIPK3(h) 2Pim-2(h) 2 Flt3(h) 5 Rsk1(h) 6 TrkA(h) 6 Rsk3(h) 7 cKit(D816H)(h) 8IRAK4(h) 12 Pim-3(h) 12 Rsk4(h) 12 MELK(h) 13 Rsk2(h) 13 CK1γ2(h) 17Flt4(h) 17 Fms(h) 17 PDGFRα(D842V)(h) 17 EGFR(T790M, L858R)(h) 20CK1γ3(h) 21 Lck(h) 21 Met(h) 22 GSK3β(h) 23 Flt3(D835Y)(h) 24 MLK1(h) 24Yes(h) 26 TAK1(h) 28 CK1γ1(h) 30 FAK(h) 30 CDK2/cyclinA(h) 31CDK1/cyclinB(h) 37 CDK9/cyclin T1(h) 37 cKit(V560G)(h) 38 Mer(h) 38ARK5(h) 39 JAK2(h) 39 PKCθ(h) 39 PKG1α(h) 40 Aurora-A(h) 43 KDR(h) 43Ret(h) 43 MST1(h) 44 Fyn(h) 49 CDK7/cyclinH/MAT1(h) 50 MSK2(h) 51EGFR(T790M)(h) 53 Mnk2(h) 54 EGFR(L858R)(h) 56 CK2(h) 58 EGFR(L861Q)(h)60 Hck(h) 61 Flt1(h) 62 LOK(h) 63 cSRC(h) 64 c-RAF(h) 66 MEK1(h) 72CK2α2(h) 73 DRAK1(h) 75 Lyn(h) 75 ErbB4(h) 77 MAPK1(h) 77 p70S6K(h) 77Snk(h) 79 MKK7β(h) 81 Fes(h) 84 PKD2(h) 86 Abl(h) 87 EphB4(h) 87 cKit(h)89 CaMKI(h) 90 DDR2(h) 90 Fer(h) 90 Ros(h) 90 ASK1(h) 92 FGFR2(h) 93PDGFRβ(h) 94 ROCK-I(h) 94 EphA5(h) 95 EphA7(h) 96 Plk1(h) 96 PDGFRα(h)97 PKA(h) 97 PRAK(h) 97 ZAP-70(h) 97 PKBα(h) 98 mTOR(h) 99 PKCα(h) 99Ron(h) 99 FGFR1(h) 100 ZIPK(h) 100 IGF-1R(h) 101 PDK1(h) 101 PAK2(h) 106SRPK1(h) 107 CHK1(h) 108 IKKα(h) 108 Tie2(h) 108 Rse(h) 109 eEF-2K(h)111 EGFR(h) 111 IR(h) 112

Estimated IC₅₀ values of compound A are as follows:

Compound Kinase IC₅₀ (nM) A Flt3(h) 104 A Pim-1(h) 1 A Pim-2(h) 6 APim-3(h) 86 A Rsk1(h) 41 A Rsk2(h) 72 A Rsk3(h) 73 A Rsk4(h) 37

The following table is the %-activity data of compound B in differentkinases at 0.5 μM of ATP.

TABLE 48 Kinase % activity HIPK3(h) −1 Flt3(D835Y)(h) 2 HIPK2(h) 2DYRK2(h) 5 Flt3(h) 7 cKit(D816H)(h) 9 Pim-1(h) 9 MELK(h) 14 DRAK1(h) 15PDGFRα(D842V)(h) 15 CDK2/cyclinA(h) 17 CDK7/cyclinH/MAT1(h) 17CDK1/cyclinB(h) 18 CDK9/cyclin T1(h) 18 ZIPK(h) 19 Rsk1(h) 21 TrkA(h) 23Lck(h) 24 GSK3β(h) 25 cKit(V560G)(h) 33 Mnk2(h) 33 PKG1α(h) 34 CK1γ3(h)38 Mer(h) 39 Rsk2(h) 40 Rsk3(h) 40 Flt4(h) 41 Rsk4(h) 41 Fms(h) 42Yes(h) 44 Pim-2(h) 46 CK2(h) 48 Fyn(h) 49 CK2α2(h) 51 EGFR(L861Q)(h) 54JAK2(h) 55 EGFR(L858R)(h) 56

 GFR(T790M, L858R)( 

59 CaMKI(h) 62 Hck(h) 62 CDK6/cyclinD3(h) 63 CK1γ2(h) 63 MSK2(h) 65Pim-3(h) 65 Flt1(h) 66 c-RAF(h) 69 CK1γ1(h) 71 Ret(h) 73 cSRC(h) 74KDR(h) 75 Lyn(h) 75 MKK7β(h) 75 EGFR(T790M)(h) 76 Plk1(h) 76 Aurora-A(h)77 ErbB4(h) 77 MLK1(h) 77 TAK1(h) 79 MST1(h) 80 IRAK4(h) 81 Snk(h) 82PKCθ(h) 83 PRAK(h) 84 MAPK1(h) 86 PKD2(h) 86 LOK(h) 87 p70S6K(h) 87Rse(h) 87 cKit(h) 88 MEK1(h) 88 PDK1(h) 88 EphA7(h) 90 IGF-1R(h) 90CHK1(h) 91 SRPK1(h) 91 Abl(h) 92 ASK1(h) 93 eEF-2K(h) 93 EphA5(h) 93FGFR1(h) 93 Tie2(h) 93 Fes(h) 94 FGFR2(h) 94 PDGFRβ(h) 94 IR(h) 95NEK2(h) 95 Ron(h) 95 Met(h) 96 PKCα(h) 97 ROCK-I(h) 97 ARK5(h) 98IKKα(h) 99 PKBα(h) 99 ALK(h) 100 PKA(h) 100 EGFR(h) 101 mTOR(h) 101Plk3(h) 102 Fer(h) 103 MAPKAP-K2(h) 104 EphB4(h) 105 PDGFRα(h) 105FAK(h) 106 ZAP-70(h) 106 PAK2(h) 108

indicates data missing or illegible when filed

The entirety of each patent, patent application, publication anddocument referenced herein hereby is incorporated by reference. Citationof the above patents, patent applications, publications and documents isnot an admission that any of the foregoing is pertinent prior art, nordoes it constitute any admission as to the contents or date of thesepublications or documents.

Modifications may be made to the foregoing without departing from thebasic aspects of the invention. Although the invention has beendescribed in substantial detail with reference to one or more specificembodiments, those of ordinary skill in the art will recognize thatchanges may be made to the embodiments specifically disclosed in thisapplication, and yet these modifications and improvements are within thescope and spirit of the invention. The invention illustrativelydescribed herein suitably may be practiced in the absence of anyelement(s) not specifically disclosed herein. Thus, for example, in eachinstance herein any of the terms “comprising”, “consisting essentiallyof”, and “consisting of” may be replaced with either of the other twoterms. Thus, the terms and expressions which have been employed are usedas terms of description and not of limitation, equivalents of thefeatures shown and described, or portions thereof, are not excluded, andit is recognized that various modifications are possible within thescope of the invention.

Representative embodiments of the invention are set forth in thefollowing aspects and illustrate but do not limit the invention.

E1. A compound of Formula IA:

wherein:

Z⁶⁰ and Z⁷⁰ are independently N or CR⁶⁰, provided at least one of themis N;

each R³⁰ and each R⁶⁰ is independently H or an optionally substitutedC1-C8 alkyl, C2-C8 heteroalkyl, C2-C8 alkenyl, C2-C8 heteroalkenyl,C2-C8 alkynyl, C2-C8 heteroalkynyl, C1-C8 acyl, C2-C8 heteroacyl, C6-C10aryl, C5-C12 heteroaryl, C7-C12 arylalkyl, or C6-C12 heteroarylalkylgroup,

-   -   or each R³⁰ and each R⁶⁰ can be halo, OR, NR₂, NROR, NRNR₂, SR,        SOR, SO₂R, SO₂NR₂, NRSO₂R, NRCONR₂, NRCSNR₂, NRC(═NR)NR₂,        NRCOOR, NRCOR, CN, COOR, CONR₂, OOCR, COR, or NO₂,    -   wherein each R is independently H or C1-C8 alkyl, C2-C8        heteroalkyl, C2-C8 alkenyl, C2-C8 heteroalkenyl, C2-C8 alkynyl,        C2-C8 heteroalkynyl, C1-C8 acyl, C2-C8 heteroacyl, C6-C₁₀ aryl,        C5-C10 heteroaryl, C7-C12 arylalkyl, or C6-C12 heteroarylalkyl,        -   and wherein two R on the same atom or on adjacent atoms can            be linked to form a 3-8 membered ring, optionally containing            one or more N, O or S;        -   and each R group, and each ring formed by linking two R            groups together, is optionally substituted with one or more            substituents selected from halo, ═O, ═N—CN, ═N—OR′, ═NR′,            OR′, NR′₂, SR′, SO₂R′, SO₂NR′₂, NR′SO₂R′, NR′CONR′₂,            NR′CSNR′₂, NR′C(═NR′)NR′₂, NR′COOR′, NR′COR′, CN, COOR′,            CONR′₂, OOCR′, COR′, and NO₂,        -   wherein each R′ is independently H, C1-C6 alkyl, C2-C6            heteroalkyl, C1-C6 acyl, C2-C6 heteroacyl, C6-C10 aryl,            C5-C10 heteroaryl, C7-12 arylalkyl, or C6-12            heteroarylalkyl, each of which is optionally substituted            with one or more groups selected from halo, C1-C4 alkyl,            C1-C4 heteroalkyl, C1-C6 acyl, C1-C6 heteroacyl, hydroxy,            amino, and ═O;        -   and wherein two R′ can be linked to form a 3-7 membered ring            optionally containing up to three heteroatoms selected from            N, O and S,        -   each R⁴⁰ is H or optionally substituted member selected from            the group consisting of C₁-C₆ alkyl, C2-C6 heteroalkyl, and            C1-C6 acyl;

each R⁵⁰ is independently an optionally substituted member selected fromthe group consisting of C₁₋₁₀ alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ heteroalkyl,C₃₋₈ carbocyclic ring, and C₃₋₈ heterocyclic ring optionally fused to anadditional optionally substituted carbocyclic or heterocyclic ring;

or R⁵⁰ can be a C₁₋₁₀ alkyl, C₂₋₁₀ alkenyl, or C₂₋₁₀ heteroalkylsubstituted with an optionally substituted C₃₋₈ carbocyclic ring or C₃₋₈heterocyclic ring;

in each —NR⁴⁰R⁵⁰, R⁴⁰ and R⁵⁰ together with N may form an optionallysubstituted 3-8 membered ring, which may optionally contain anadditional heteroatom selected from N, O and S as a ring member; and

each R^(3P) represents a polar substituent;

or a pharmaceutically acceptable salt thereof.

E2. The compound of embodiment E1, wherein Z⁶⁰ is N and Z⁷⁰ is CH.

E3. The compound of embodiment E1, wherein Z⁷⁰ is N and Z⁶⁰ is CH.

E4. The compound of embodiment E1, E2 or E3, wherein each R⁶⁰ and R⁴⁰ isH.

E5. The compound of any one of embodiments E1 to E4, wherein R^(3P) isan optionally substituted imidazole or triazole ring.

E6. The compound of any one of embodiments E1 to E5, wherein R⁵⁰ isunsubstituted phenyl or phenyl substituted with 1-3 substituentsselected from halo, cyano, CF₃, —OCF₃, COOR⁴⁰, and SO₂NR⁴⁰R⁵⁰, and oneor more of these substituents can be an optionally substituted groupselected from C1-C6 alkyl, C1-C6 alkoxy, C2-C6 alkenyl, and C2-C6alkynyl.

E7. The compound of embodiment E1, which is a compound of Formula IB:

or a pharmaceutically acceptable salt thereof,

wherein R³⁰ is as defined for embodiment 1,

and R^(3P) is an optionally substituted imidazole or triazole ring;

and each Φ independently represents an optionally substituted phenyl.

E8. The compound of embodiment E1, which is a compound of Formula IC:

or a pharmaceutically acceptable salt thereof,

wherein R³⁰ is as defined for embodiment 1,

and R^(3P) is an optionally substituted imidazole or triazole ring;

and each Φ independently represents an optionally substituted phenyl.

E9. The compound of embodiment E7 or E8, wherein Φ is unsubstitutedphenyl or phenyl substituted with 1-3 substituents selected from halo,cyano, CF₃, —OCF₃, COOR⁴⁰, and SO₂NR⁴⁰R⁵⁰, and one or more of thesesubstituents can be an optionally substituted group selected from C1-C6alkyl, C1-C6 alkoxy, C2-C6 alkenyl, and C2-C6 alkynyl.

E10. The compound of any one of embodiments E1 to E5, wherein R⁵⁰ is anoptionally substituted C₃₋₈ carbocyclic or C₃₋₈ heterocyclic ring, eachof which may be optionally fused to an additional optionally substitutedcarbocyclic or heterocyclic ring.

E11. The compound of embodiment E10, wherein said optionally substitutedC₃₋₈ carbocyclic or C₃₋₈ heterocyclic ring is an optionally substitutedaromatic or heteroaromatic ring.

E12. A compound selected from the group consisting of:

or a pharmaceutically acceptable salt thereof.

E13. A compound of Formula L:

or a pharmaceutically acceptable salt thereof;

wherein:

Z⁶⁰ and Z⁷⁰ are independently N or CR⁶⁰, provided at least one of themis N;

each R³⁰ and each R⁶⁰ is independently H or an optionally substitutedC1-C8 alkyl, C2-C8 heteroalkyl, C2-C8 alkenyl, C2-C8 heteroalkenyl,C2-C8 alkynyl, C2-C8 heteroalkynyl, C1-C8 acyl, C2-C8 heteroacyl, C6-C10aryl, C5-C12 heteroaryl, C7-C12 arylalkyl, or C6-C12 heteroarylalkylgroup,

or each R³⁰ and each R⁶⁰ can be halo, OR, NR₂, NROR, NRNR₂, SR, SOR,SO₂R, SO₂NR₂, NRSO₂R, NRCONR₂, NRCSNR₂, NRC(═NR)NR₂, NRCOOR, NRCOR, CN,COOR, CONR₂, OOCR, COR, or NO₂,

wherein each R is independently H or C1-C8 alkyl, C2-C8 heteroalkyl,C2-C8 alkenyl, C2-C8 heteroalkenyl, C2-C8 alkynyl, C2-C8 heteroalkynyl,C1-C8 acyl, C2-C8 heteroacyl, C6-C10 aryl, C5-C10 heteroaryl, C7-C12arylalkyl, or C6-C12 heteroarylalkyl,

and wherein two R on the same atom or on adjacent atoms can be linked toform a 3-8 membered ring, optionally containing one or more N, O or S;

and each R group, and each ring formed by linking two R groups together,is optionally substituted with one or more substituents selected fromhalo, ═O, ═N—CN, ═N—OR′, ═NR′, OR′, NR′₂, SR′, SO₂R′, SO₂NR′₂, NR′SO₂R′,NR′CONR′₂, NR′CSNR′₂, NR′C(═NR′)NR′₂, NR′COOR′, NR′COR′, CN, COOR′,CONR′₂, OOCR′, COR′, and NO₂,

wherein each R′ is independently H, C1-C6 alkyl, C2-C6 heteroalkyl,C1-C6 acyl, C2-C6 heteroacyl, C6-C10 aryl, C5-C10 heteroaryl, C7-12arylalkyl, or C6-12 heteroarylalkyl, each of which is optionallysubstituted with one or more groups selected from halo, C1-C4 alkyl,C1-C4 heteroalkyl, C1-C6 acyl, C1-C6 heteroacyl, hydroxy, amino, and ═O;

and wherein two R′ can be linked to form a 3-7 membered ring optionallycontaining up to three heteroatoms selected from N, O and S,

each R^(3P) represents a polar substituent; and

each W represents an optionally substituted aryl, an optionallysubstituted heteroaryl, or an optionally substituted C₃₋₈ cycloalkylring.

E14. The compound of embodiment E13, which is a compound of Formula L-Aor Formula L-B:

-   -   or a pharmaceutically acceptable salt thereof.

E15. The compound of embodiment E13 or E14, wherein R^(3P) is anoptionally substituted imidazole or triazole ring.

E16. A pharmaceutical composition comprising a compound of any one ofembodiments E1 to E12 and a pharmaceutically acceptable excipient.

E17. A pharmaceutical composition comprising a compound of embodimentE13, E14 or E15 and a pharmaceutically acceptable excipient.

E18. A method for inhibiting cell proliferation, which comprisescontacting cells with a compound or composition according to any one ofembodiments E1 to E17, in an amount effective to inhibit proliferationof the cells.

E19. The method of embodiment E18, wherein the cells are in a cancercell line.

E20. The method of embodiment E19, wherein the cancer cell line is abreast cancer, prostate cancer, pancreatic cancer, lung cancer,hemopoietic cancer, colorectal cancer, skin cancer, ovary cancer cellline.

E21. The method of embodiment E18 or E19, wherein the cells are in atumor in a subject.

E22. The method of any one of embodiments E18 to E21, wherein contactingcells with a compound having a structure of any one of embodiments E1 toE11 induces cell apoptosis.

E23. A method for treating a condition related to aberrant cellproliferation, which comprises administering a compound or compositionaccording to any one of embodiments E1 to E17 to a subject in needthereof in an amount effective to treat the cell proliferativecondition.

E24. The method of embodiment E23, wherein the cell proliferativecondition is a tumor-associated cancer.

E25. The method of embodiment E24, wherein the cancer is of thecolorectum, breast, lung, liver, pancreas, lymph node, colon, prostate,brain, head and neck, skin, liver, kidney, blood and heart.

E26. The method of embodiment E23, wherein the cell proliferativecondition is a non-tumor cancer.

E27. The method of embodiment E26, wherein the non-tumor cancer is ahematopoietic cancer.

E28. The method of embodiment E27, wherein the hematopoietic cancer isacute myelogenous leukemia.

E29. The method of embodiment E28, wherein the leukemia is refractoryAML or wherein the AML is associated with a mutated Flt3.

E30. A method for treating pain or inflammation in a subject, whichcomprises administering a compound or composition according to any oneof embodiments E1 to E11 to a subject in need thereof in an amounteffective to treat the pain or the inflammation.

LENGTHY TABLES The patent application contains a lengthy table section.A copy of the table is available in electronic form from the USPTO website(http://seqdata.uspto.gov/?pageRequest=docDetail&DocID=US20120208792A1).An electronic copy of the table will also be available from the USPTOupon request and payment of the fee set forth in 37 CFR 1.19(b)(3).

1. (canceled)
 2. The method of claim 23, wherein Z⁶⁰ is N and Z⁷⁰ is CH.3. The method of claim 23, wherein Z⁷⁰ is N and Z⁶⁰ is CH.
 4. The methodof claim 23, wherein each R⁶⁰ and R⁴⁰ is H.
 5. The method of claim 23,wherein R^(3P) is an optionally substituted imidazole or triazole ring.6. The method of claim 23, wherein R⁵⁰ is unsubstituted phenyl or phenylsubstituted with 1-3 substituents selected from halo, cyano, CF₃, —OCF₃,COOR⁴⁰, and SO₂NR⁴⁰R⁵⁰, and one or more of these substituents can be anoptionally substituted group selected from C1-C6 alkyl, C1-C6 alkoxy,C2-C6 alkenyl, and C2-C6 alkynyl.
 7. The method of claim 23, wherein thecompound of Formula IA is represented by Formula IB:

or a pharmaceutically acceptable salt thereof, wherein R³⁰ is as definedfor claim 23, and R^(3P) is an optionally substituted imidazole ortriazole ring; and each Φ independently represents an optionallysubstituted phenyl.
 8. The method of claim 23, wherein the compound ofFormula IA is represented by Formula IC:

or a pharmaceutically acceptable salt thereof, wherein R³⁰ is as definedfor claim 23, and R^(3P) is an optionally substituted imidazole ortriazole ring; and each Φ independently represents an optionallysubstituted phenyl.
 9. The method of claim 8, wherein Φ is unsubstitutedphenyl or phenyl substituted with 1-3 substituents selected from halo,cyano, CF₃, —OCF₃, COOR⁴⁰, and SO₂NR⁴⁰R⁵⁰, and one or more of thesesubstituents can be an optionally substituted group selected from C1-C6alkyl, C1-C6 alkoxy, C2-C6 alkenyl, and C2-C6 alkynyl.
 10. The method ofclaim 23, wherein R⁵⁰ is an optionally substituted C₃₋₈ carbocyclic orC₃₋₈ heterocyclic ring, each of which may be optionally fused to anadditional optionally substituted carbocyclic or heterocyclic ring. 11.The method of claim 10, wherein said optionally substituted C₃₋₈carbocyclic or C₃₋₈ heterocyclic ring is an optionally substitutedaromatic or heteroaromatic ring.
 12. The method of claim 23, wherein thecompound is selected from the group consisting of:

or a pharmaceutically acceptable salt thereof. 13-17. (canceled)
 18. Amethod for inhibiting cell proliferation, which comprises contactingcells with a compound having a structure of Formula IA, or apharmaceutically acceptable salt thereof, in an amount effective toinhibit proliferation of the cells,

wherein: Z⁶⁰ and Z⁷⁰ are independently N or CR⁶⁰, provided at least oneof them is N; each R³⁰ and each R⁶⁰ is independently H or an optionallysubstituted C1-C8 alkyl, C2-C8 heteroalkyl, C2-C8 alkenyl, C2-C8heteroalkenyl, C2-C8 alkynyl, C2-C8 heteroalkynyl, C1-C8 acyl, C2-C8heteroacyl, C6-C10 aryl, C5-C12 heteroaryl, C7-C12 arylalkyl, or C6-C12heteroarylalkyl group, or each R³⁰ and each R⁶⁰ can be halo, OR, NR₂,NROR, NRNR₂, SR, SOR, SO₂R, SO₂NR₂, NRSO₂R, NRCONR₂, NRCSNR₂,NRC(═NR)NR₂, NRCOOR, NRCOR, CN, COOR, CONR₂, OOCR, COR, or NO₂, whereineach R is independently H or C1-C8 alkyl, C2-C8 heteroalkyl, C2-C8alkenyl, C2-C8 heteroalkenyl, C2-C8 alkynyl, C2-C8 heteroalkynyl, C1-C8acyl, C2-C8 heteroacyl, C6-C10 aryl, C5-C10 heteroaryl, C7-C12arylalkyl, or C6-C12 heteroarylalkyl, and wherein two R on the same atomor on adjacent atoms can be linked to form a 3-8 membered ring,optionally containing one or more N, O or S; and each R group, and eachring formed by linking two R groups together, is optionally substitutedwith one or more substituents selected from halo, ═O, ═N—CN, ═N—OR′,═NR′, OR′, NR′₂, SR′, SO₇R′, SO₂NR′₂, NR′SO₂R′, NR′CONR′₂, NR′CSNR′₂,NR′C(═NR′)NR′₂, NR′COOR′, NR′COR′, CN, COOR′, CONR′₂, OOCR′, COR′, andNO₂, wherein each R′ is independently H, C1-C6 alkyl, C2-C6 heteroalkyl,C1-C6 acyl, C2-C6 heteroacyl, C6-C10 aryl, C5-C10 heteroaryl, C7-12arylalkyl, or C6-12 heteroarylalkyl, each of which is optionallysubstituted with one or more groups selected from halo, C1-C4 alkyl,C1-C4 heteroalkyl, C1-C6 acyl, C1-C6 heteroacyl, hydroxy, amino, and ═O;and wherein two R′ can be linked to form a 3-7 membered ring optionallycontaining up to three heteroatoms selected from N, O and S, each R⁴⁰ isH or optionally substituted member selected from the group consisting ofC₁-C₆ alkyl, C2-C6 heteroalkyl, and C1-C6 acyl; each R⁵⁰ isindependently an optionally substituted member selected from the groupconsisting of C₁₋₁₀ alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ heteroalkyl, C₃₋₈carbocyclic ring, and C₃₋₈ heterocyclic ring optionally fused to anadditional optionally substituted carbocyclic or heterocyclic ring; orR⁵⁰ can be a C₁₋₁₀ alkyl, C₂₋₁₀ alkenyl, or C₂₋₁₀ heteroalkylsubstituted with an optionally substituted C₃₋₈ carbocyclic ring or C₃₋₈heterocyclic ring; in each —NR⁴⁰R⁵⁰, R⁴⁰ and R⁵⁰ together with N mayform an optionally substituted 3-8 membered ring, which may optionallycontain an additional heteroatom selected from N, O and S as a ringmember; and each R^(3P) represents a polar substituent.
 19. The methodof claim 18, wherein the cells are in a cancer cell line.
 20. The methodof claim 19, wherein the cancer cell line is a breast cancer, prostatecancer, pancreatic cancer, lung cancer, hemopoietic cancer, colorectalcancer, skin cancer, ovary cancer cell line.
 21. The method of claim 18,wherein the cells are in a tumor in a subject.
 22. The method of claim18, wherein contacting cells with a compound having a structure ofFormula IA induces cell apoptosis.
 23. A method for treating a conditionrelated to aberrant cell proliferation, which comprises administering acompound having a structure of Formula IA, or a pharmaceuticallyacceptable salt thereof, to a subject in need thereof in an amounteffective to treat the cell proliferative condition,

wherein: Z⁶⁰ and Z⁷⁰ are independently N or CR⁶⁰, provided at least oneof them is N; each R³⁰ and each R⁶⁰ is independently H or an optionallysubstituted C1-C8 alkyl, C2-C8 heteroalkyl, C2-C8 alkenyl, C2-C8heteroalkenyl, C2-C8 alkynyl, C2-C8 heteroalkynyl, C1-C8 acyl, C2-C8heteroacyl, C6-C10 aryl, C5-C12 heteroaryl, C7-C12 arylalkyl, or C6-C12heteroarylalkyl group, or each R³⁰ and each R⁶⁰ can be halo, OR, NR₂,NROR, NRNR₂, SR, SOR, SO₂R, SO₂NR₂, NRSO₂R, NRCONR₂, NRCSNR₂,NRC(═NR)NR₂, NRCOOR, NRCOR, CN, COOR, CONR₂, OOCR, COR, or NO₂, whereineach R is independently H or C1-C8 alkyl, C2-C8 heteroalkyl, C2-C8alkenyl, C2-C8 heteroalkenyl, C2-C8 alkynyl, C2-C8 heteroalkynyl, C1-C8acyl, C2-C8 heteroacyl, C6-C10 aryl, C5-C10 heteroaryl, C7-C12arylalkyl, or C6-C12 heteroarylalkyl, and wherein two R on the same atomor on adjacent atoms can be linked to form a 3-8 membered ring,optionally containing one or more N, O or S; and each R group, and eachring formed by linking two R groups together, is optionally substitutedwith one or more substituents selected from halo, ═O, ═N—CN, ═NR′, OR′,NR′₂, SR′, SO₂R′, SO₂NR′₂, NR′SO₂R′, NR′CONR′₂, NR′CSNR′₂,NR′C(═NR′)NR′₂, NR′COOR′, NR′COR′, CN, COOR′, CONR′₂, OOCR′, COR′, andNO₂, wherein each R′ is independently H, C1-C6 alkyl, C2-C6 heteroalkyl,C1-C6 acyl, C2-C6 heteroacyl, C6-C10 aryl, C5-C10 heteroaryl, C7-12arylalkyl, or C6-12 heteroarylalkyl, each of which is optionallysubstituted with one or more groups selected from halo, C1-C4 alkyl,C1-C4 heteroalkyl, C1-C6 acyl, C1-C6 heteroacyl, hydroxy, amino, and ═O;and wherein two R′ can be linked to form a 3-7 membered ring optionallycontaining up to three heteroatoms selected from N, O and S, each R⁴⁰ isH or optionally substituted member selected from the group consisting ofC₁-C₆ alkyl, C2-C6 heteroalkyl, and C1-C6 acyl; each R⁵⁰ isindependently an optionally substituted member selected from the groupconsisting of C₁₋₁₀ alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ heteroalkyl, C₃₋₈carbocyclic ring, and C₃₋₈ heterocyclic ring optionally fused to anadditional optionally substituted carbocyclic or heterocyclic ring; orR⁵⁰ can be a C₁₋₁₀ alkyl, C₂₋₁₀ alkenyl, or C₂₋₁₀ heteroalkylsubstituted with an optionally substituted C₃₋₈ carbocyclic ring or C₃₋₈heterocyclic ring; in each —NR⁴⁰R⁵⁰, R⁴⁰ and R⁵⁰ together with N mayform an optionally substituted 3-8 membered ring, which may optionallycontain an additional heteroatom selected from N, O and S as a ringmember; and each R^(3P) represents a polar substituent.
 24. The methodof claim 23, wherein the cell proliferative condition is atumor-associated cancer.
 25. The method of claim 24, wherein the canceris of the colorectum, breast, lung, liver, pancreas, lymph node, colon,prostate, brain, head and neck, skin, liver, kidney, blood and heart.26. The method of claim 23, wherein the cell proliferative condition isa non-tumor cancer.
 27. The method of claim 26, wherein the non-tumorcancer is a hematopoietic cancer.
 28. The method of claim 27, whereinthe hematopoietic cancer is acute myelogenous leukemia.
 29. The methodof claim 28, wherein the leukemia is refractory AML or wherein the AMLis associated with a mutated Flt3.
 30. A method for treating pain orinflammation in a subject, which comprises administering a compound ofFormula IA, or a pharmaceutically acceptable salt thereof, to a subjectin need thereof in an amount effective to treat the pain or theinflammation,

wherein: Z⁶⁰ and Z⁷⁰ are independently N or CR⁶⁰, provided at least oneof them is N; each R³⁰ and each R⁶⁰ is independently H or an optionallysubstituted C1-C8 alkyl, C2-C8 heteroalkyl, C2-C8 alkenyl, C2-C8heteroalkenyl, C2-C8 alkynyl, C2-C8 heteroalkynyl, C1-C8 acyl, C2-C8heteroacyl, C6-C10 aryl, C5-C12 heteroaryl, C7-C12 arylalkyl, or C6-C12heteroarylalkyl group, or each R³⁰ and each R⁶⁰ can be halo, OR, NR₂,NROR, NRNR₂, SR, SOR, SO₂R, SO₂NR₂, NRSO₂R, NRCONR₂, NRCSNR₂,NRC(═NR)NR₂, NRCOOR, NRCOR, CN, COOR, CONR₂, OOCR, COR, or NO₂, whereineach R is independently H or C1-C8 alkyl, C2-C8 heteroalkyl, C2-C8alkenyl, C2-C8 heteroalkenyl, C2-C8 alkynyl, C2-C8 heteroalkynyl, C1-C8acyl, C2-C8 heteroacyl, C6-C10 aryl, C5-C10 heteroaryl, C7-C12arylalkyl, or C6-C12 heteroarylalkyl, and wherein two R on the same atomor on adjacent atoms can be linked to form a 3-8 membered ring,optionally containing one or more N, O or S; and each R group, and eachring formed by linking two R groups together, is optionally substitutedwith one or more substituents selected from halo, ═O, ═N—CN, ═N—OR′,═NR′, OR′, NR′₂, SR′, SO₂R′, SO₂NR′₂, NR′SO₂R′, NR′CONR′₂, NR′CSNR′₂,NR′C(═NR′)NR′₂, NR′COOR′, NR′COR′, CN, COOR′, CONR′₂, OOCR′, COR′, andNO₂, wherein each R′ is independently H, C1-C6 alkyl, C2-C6 heteroalkyl,C1-C6 acyl, C2-C6 heteroacyl, C6-C10 aryl, C5-C10 heteroaryl, C7-12arylalkyl, or C6-12 heteroarylalkyl, each of which is optionallysubstituted with one or more groups selected from halo, C1-C4 alkyl,C1-C4 heteroalkyl, C1-C6 acyl, C1-C6 heteroacyl, hydroxy, amino, and ═O;and wherein two R′ can be linked to form a 3-7 membered ring optionallycontaining up to three heteroatoms selected from N, O and S, each R₄₀ isH or optionally substituted member selected from the group consisting ofC₁-C₆ alkyl, C2-C6 heteroalkyl, and C1-C6 acyl; each R⁵⁰ isindependently an optionally substituted member selected from the groupconsisting of C₁₋₁₀ alkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ heteroalkyl, C₃₋₈carbocyclic ring, and C₃₋₈ heterocyclic ring optionally fused to anadditional optionally substituted carbocyclic or heterocyclic ring; orR⁵⁰ can be a C₁₋₁₀ alkyl, C₂₋₁₀ alkenyl, or C₂₋₁₀ heteroalkylsubstituted with an optionally substituted C₃₋₈ carbocyclic ring or C₃₋₈heterocyclic ring; in each —NR⁴⁰R⁵⁰, R⁴⁰ and R⁵⁰ together with N mayform an optionally substituted 3-8 membered ring, which may optionallycontain an additional heteroatom selected from N, O and S as a ringmember; and each R^(3P) represents a polar substituent.